Compositions and methods for modulating farnesoid x receptors

ABSTRACT

The present invention relates to compounds of Formula I, 
     
       
         
         
             
             
         
       
     
     a stereoisomer, enantiomer, a pharmaceutically acceptable salt or an amino acid conjugate thereof; wherein variables are as defined herein; and their pharmaceutical compositions, which are useful as modulators of the activity of Farnesoid X receptors (FXR).

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/034,280 filed May 4, 2016, which is a 371 U.S. national phaseapplication of international application number PCT/US2014/063948 filedNov. 4, 2014 and claims the benefit of U.S. provisional application Ser.No. 61/900,013, filed Nov. 5, 2013. Each of these applications isincorporated by reference herein in its entirety.

The instant application contains a Sequence Listing, which has beenfiled electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on May 18, 2017, isnamed PAT055829-US-CNT02 Sequence Listing_ST25.txt and is 929 bytes insize.

FIELD OF THE INVENTION

The present invention relates to compositions and methods for modulatingthe activity of farnesoid X receptors (FXRs).

BACKGROUND OF THE INVENTION

The farnesoid X receptor (FXR) is a member of the nuclear hormonereceptor superfamily and is primarily expressed in the liver, kidney andintestine (see, e.g., Seol et al. (1995) Mol. Endocrinol. 9:72-85 andForman et al. (1995) Cell 81:687-693). It functions as a heterodimerwith the retinoid X receptor (RXR) and binds to response elements in thepromoters of target genes to regulate gene transcription. The FXR-RXRheterodimer binds with highest affinity to an inverted repeat-1 (IR-1)response element, in which consensus receptor-binding hexamers areseparated by one nucleotide. FXR is part of an interrelated process, inthat FXR is activated by bile acids (the end product of cholesterolmetabolism) (see, e.g., Makishima et al. (1999) Science 284: 1362-1365,Parks et al. (1999) Science 284:1365-1368, Wang et al. (1999) Mol. Cell.3:543-553), which serve to inhibit cholesterol catabolism. See also,Urizar et al. (2000) J. Biol. Chem. 275:39313-39317.

FXR is a key regulator of cholesterol homeostasis, triglyceridesynthesis and lipogenesis. (Crawley, Expert Opinion Ther. Patents(2010), 20(8): 1047-1057). In addition to the treatment of dyslipidemia,multiple indications for FXR have been described, including treatment ofliver disease, diabetes, vitamin D-related diseases, drug-induced sideeffects and hepatitis. (Crawley, supra). While advances have been madein the development of novel FXR agonists, there remains significant roomfor improvement.

SUMMARY OF THE INVENTION

The present invention relates to compositions and methods for modulatingthe activity of farnesoid X receptors (FXRs). For example, the presentinvention provides novel compounds that are agonists or partial agonistsof FXR, and are useful as pharmaceuticals to treat FXR-mediatedconditions.

In one aspect, the compounds of the invention are defined by Formula(I):

or a pharmaceutical acceptable salt thereof, wherein,

R⁰ is Ring A or C₁₋₆ alkyl;

Ring A is aryl; 5-10 membered heteroaryl comprising 1-3 N, O or Sheteroatoms; 4-6 membered heterocycle comprising 1-2 N, O or Sheteroatoms; or C₃₋₇ cycloalkyl; and said Ring A is unsubstituted orsubstituted by 1-2 substituents independently represented by R²;

Ring B is aryl; 5-10 membered heteroaryl comprising 1-3 N, O or Sheteroatoms; 4-6 membered heterocycle comprising 1-2 N, O or Sheteroatoms; or C₃₋₇ cycloalkyl; and said Ring B is unsubstituted orsubstituted by 1-2 substituents independently represented by R²;

X is —(CR⁴R⁵)— or —C(O)—;

Y is —O—, —(CR⁴R⁵)—, *—O(CR⁴R⁵)— or —NR—, wherein “*” indicates thepoint of attachment of Y to the ring containing the Z ring atoms;

Z¹, Z², Z³, and Z⁴ are each independently —CR³— or —N—;

L¹ is *¹—(CR⁴R⁵)₁₋₂— or *¹—(CR⁴R⁵)—C(O)—NR—, wherein “*¹” indicates thepoint of attachment of L¹ to N;

L² is *²—(CR⁴R⁵)₁₋₂—, *²—(CR⁴R⁵)—C(O)—, *²—(CR⁴R⁵)—C(O)—NR—,*²—(CR⁴R⁵)₂—O—, *²—(CR⁴R⁵)₂—NR—, *²—(CR⁴R⁵)₂—SO₂—, *²—(CR⁴R⁵)₂—NR—C(O)—,or *²—(CR⁴R⁵)—C(O)—NR—(CR⁴R⁵)—; wherein “*²” indicates the point ofattachment of L² to N;

L³ is —(CR⁴R⁵)— or —C(O)—;

each R² is independently halo, hydroxyl, C₁₋₆ alkyl, or halo-substitutedC₁₋₆ alkyl;

each R³ is independently hydrogen, halo, or C₁₋₆ alkyl; and

R, R⁴ and R⁵ are independently hydrogen or C₁₋₆ alkyl.

In one embodiment, the compounds of the invention are defined by Formula(II):

or a pharmaceutically acceptable salt thereof; wherein,

-   -   R⁰ is Ring A or C₁₋₆ alkyl; wherein Ring A is phenyl, pyridyl or        cyclopropyl, each of which is unsubstituted or substituted by        1-2 substituents independently represented by R²;    -   Ring B is selected from phenyl, pyridyl, 1H-indolyl, and C₃₋₇        cycloalkyl, each of which is unsubstituted or substituted by 1-2        substituents independently represented by R²;    -   X is —(CR⁴R⁵)—;    -   Y is —O—, —(CR⁴R⁵)— or *—O(CR⁴R⁵)—, wherein “*” indicates the        point of attachment of Y to the ring containing the Z ring        atoms;    -   Z¹, Z², Z³, and Z⁴ are each independently —CR³— or —N—;    -   L¹ is *¹—(CR⁴R⁵)₁₋₂— wherein “*¹” indicates the point of        attachment of L¹ to N;    -   L² is *²—(CR⁴R⁵)₁₋₂—, *²—(CR⁴R⁵)—C(O)—NR—, *²—(CR⁴R⁵)₂—O—,        *²—(CR⁴R⁵)₂—NR— or *²—(CR⁴R⁵)—C(O)—NR—(CR⁴R⁵)—; where “*²”        indicates the point of attachment of L² to N;    -   each R² is independently halo, C₁₋₆ alkyl, or halo-substituted        C₁₋₆ alkyl;    -   each R³ is independently hydrogen, halo, or C₁₋₆ alkyl; and    -   R, R⁴ and R⁵ are independently hydrogen or C₁₋₆ alkyl.

In another embodiment, the compounds of the invention are represented byFormula (III):

or a pharmaceutically acceptable salt thereof; wherein

-   -   Ring A is phenyl or pyridyl, each of which is unsubstituted or        substituted by 1-2 substituents independently represented by R²;    -   Ring B is selected from phenyl, pyridyl, 1H-indolyl, and        cyclopentyl, each of which is unsubstituted or substituted by        1-2 substituents independently represented by R²;    -   L¹ is —(CR⁴R⁵)—;    -   L² is selected from —(CH₂)—, *²—CH₂C(O)NH—, *²—CH(CH₃)C(O)NH—,        *²—CH₂C(O)NHCH₂—, *²—(CH₂)₂O—, and *²—(CH₂)₂NH—; wherein “*²”        indicates the point of attachment of L² to N;    -   X is CH₂;    -   Y is selected from —O—, —CH₂—, —C(CH₃)₂— and *—O—CH₂—, wherein        “*” indicates the point of attachment of Y to the ring        containing the Z ring atoms;    -   Z¹ is CR³ or N;    -   Z² is CR³;    -   Z³ is CR³;    -   Z⁴ is CR³ or N;    -   each R² is independently selected from halo, methyl, and        trifluoromethyl;    -   each R³ is independently hydrogen, halo, or C₁₋₆ alkyl; and    -   each of R⁴ and R⁵ is independently hydrogen or methyl.

In yet another embodiment, the compounds of the invention arerepresented by Formula (IV):

or a pharmaceutically acceptable salt thereof; wherein

-   -   each phenyl ring is optionally further substituted by 1-2        substituents independently represented by R², wherein R² is        fluoro or methyl;    -   L² is selected from —CH₂—, *²—CH₂CH₂NH—, *²—CH₂CH₂O—, and        *²—CH₂C(O)NH—, wherein “*²” indicates the point of attachment of        L² to N;    -   Z² is selected from CH, CF, CCl, and CCH₃; and    -   Z³ is selected from CH, CF, CCl, and CCH₃.

The present invention also provides a compound represented by Formula(V):

-   -   wherein each phenyl ring is optionally further substituted by        1-2 substituents independently represented by R², wherein R² is        fluoro or methyl;    -   L² is selected from —CH₂—, *²—CH₂CH₂NH—, *²—CH₂CH₂O—, and        *²—CH₂C(O)NH—, wherein “*²” indicates the point of attachment of        L² to N;    -   Z² is selected from CH, CF, CCl, and CCH₃;    -   Z³ is selected from CH, CF, CCl, and CCH₃; and    -   R⁶ is C₁₋₆ alkyl.

The compounds of Formula I, II, III, IV and V, and theirpharmaceutically acceptable salts exhibit valuable pharmacologicalproperties when tested in vitro in cell-free kinase assays and incellular assays, and are therefore useful as pharmaceuticals.

In one aspect, the invention relates to methods for modulating FXR in acell, comprising contacting the cell with an effective amount of acompound of Formula I, II, III, IV or V, or a pharmaceuticallyacceptable salt thereof; and optionally in combination with a secondtherapeutic agent.

In another aspect, the invention relates to methods to treat, ameliorateor prevent a FXR-mediated disorder in a subject suffering there from,comprising administering to the subject a therapeutically effectiveamount of a compound of Formula I, II, III, IV or V, or apharmaceutically acceptable salt thereof; and optionally in combinationwith a second therapeutic agent. The present invention also provides forthe use of a compound of Formula I, II, III, IV or V, or apharmaceutically acceptable salt thereof; and optionally in combinationwith a second therapeutic agent, in the manufacture of a medicament fortreating a FXR-mediated disorder. In yet another aspect, the presentinvention relates to a combination comprising a therapeuticallyeffective amount of a compound of Formula I, II, III, IV or V, or apharmaceutically acceptable salt thereof, and a second therapeuticagent. Where a second therapeutic agent is used, the second therapeuticagent may also be useful in the treatment of a FXR-mediated disorder.

In one embodiment, the compounds (alone or in combination with a secondtherapeutic agent) are useful for treating a liver disease or agastrointestinal disease, including but not limited to liver diseasesselected from intrahepatic cholestasis, estrogen-induced cholestasis,drug-induced cholestasis, cholestasis of pregnancy, parenteralnutrition-associated cholestasis, progressive familiar cholestasis(PFIC), Alagille syndrome, primary biliary cirrhosis (PBC), primarysclerosing cholangitis, ductopenic liver transplant rejection, livertransplant associated graft versus host disease, cystic fibrosis liverdisease, non-alcoholic fatty liver disease (NAFLD), non-alcoholicsteatohepatitis (NASH), alcoholic liver disease, and parenteralnutrition-associated liver disease; and gastrointestinal diseasesselected from bile acid malabsorption (including primary bile aciddiarrhea and secondary bile acid diarrhea), bile reflux gastritis, andinflammatory bowel disease such as Crohn's disease, ulcerative colitis,collagenous colitis, lymphocytic colitis, diversion colitis,indeterminate colitis and Behçet's disease.

DETAILED DESCRIPTION OF THE INVENTION Definitions

For purposes of interpreting this specification, the followingdefinitions will apply and whenever appropriate, terms used in thesingular will also include the plural and vice versa.

As used herein, “C₁₋₆ alkyl” denotes an alkyl radical having from 1 upto 6, in some cases from 1 up to 4 carbon atoms, the radicals beingeither linear or branched with single or multiple branching. Forexample, “C₁₋₆ alkyl” includes but is not limited n-butyl, sec-butyl,isobutyl, tert-butyl; propyl, such as n-propyl, 2-methylpropyl orisopropyl; ethyl or methyl.

As used herein, the term “alkylene” refers to divalent alkyl group asdefined herein above having 1 to 4 carbon atoms. Representative examplesof alkylene include, but are not limited to, methylene, ethylene,n-propylene, iso-propylene, n-butylene, sec-butylene, iso-butylene,tert-butylene, and the like.

As used herein, “C₃₋₇ cycloalkyl” refers to a non-aromatic saturated orpartially unsaturated monocyclic, bicyclic, bridged or spirocyclichydrocarbon groups of 3-7 carbon ring atoms. Exemplary monocyclichydrocarbon groups include, but are not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl and cyclohexenyl andthe like.

As used herein, “halogen” or “halo” refers to fluoro, chloro, bromo, andiodo; and more particularly, fluoro or chloro.

As used herein, “halo C₁₋₆ alkyl” or “halo-substituted C₁₋₆ alkyl”refers to an alkyl radical, as defined above that is substituted by oneor more halo radicals, as defined above, and is particularly fluoro C₁₋₆alkyl, more particularly trifluoromethyl.

As used herein, a “stereoisomer” refers to a compound made up of thesame atoms bonded by the same bonds but having differentthree-dimensional structures, which are not interchangeable. The presentinvention contemplates various stereoisomers and mixtures thereof andincludes “enantiomers”, which refers to two stereoisomers whosemolecules are non-superimposable mirror images of one another.

As used herein, the term “amino acid conjugate” refers to conjugates ofthe compound of Formula I, II, III, IV or V with any suitable aminoacid. Preferably, such suitable amino acid conjugates of the compound ofFormula I, II, III, IV or V will have the added advantage of enhancedintegrity in bile or intestinal fluids. Suitable amino acids include butare not limited to glycine, taurine and acyl glucuronide. Thus, thepresent invention encompasses the glycine, taurine and acyl glucuronideconjugates of the compound of Formula I, II, III, IV or V.

As used herein, the term “pharmaceutically acceptable carrier” includesany and all solvents, dispersion media, coatings, surfactants,antioxidants, preservatives (e.g., antibacterial agents, antifungalagents), isotonic agents, absorption delaying agents, salts,preservatives, drugs, drug stabilizers, binders, excipients,disintegration agents, lubricants, sweetening agents, flavoring agents,dyes, and the like and combinations thereof, as would be known to thoseskilled in the art (see, for example, Remington's PharmaceuticalSciences, 18th Ed. Mack Printing Company, 1990, pp. 1289-1329). Exceptinsofar as any conventional carrier is incompatible with the activeingredient, its use in the therapeutic or pharmaceutical compositions iscontemplated.

As used herein, the term “therapeutically effective amount” refers to anamount of the compound of formula (I) which is sufficient to achieve thestated effect. Accordingly, a therapeutically effective amount of acompound of formula (I) used in for the treatment of a conditionmediated by FXR will be an amount sufficient for the treatment of thecondition mediated by FXR.

As used herein, the term “subject” refers to an animal. Typically theanimal is a mammal. A subject also refers to for example, primates(e.g., humans, male or female), cows, sheep, goats, horses, dogs, cats,rabbits, rats, mice, fish, birds and the like. In certain embodiments,the subject is a primate. In yet other embodiments, the subject is ahuman.

As used herein, the term “treat”, “treating” or “treatment” of anydisease or disorder refers in one embodiment, to ameliorating thedisease or disorder (i.e., slowing or arresting or reducing thedevelopment of the disease or at least one of the clinical symptomsthereof). In another embodiment “treat”, “treating” or “treatment”refers to alleviating or ameliorating at least one physical parameterincluding those which may not be discernible by the patient. In yetanother embodiment, “treat”, “treating” or “treatment” refers tomodulating the disease or disorder, either physically, (e.g.,stabilization of a discernible symptom), physiologically, (e.g.,stabilization of a physical parameter), or both. In yet anotherembodiment, “treat”, “treating” or “treatment” refers to preventing ordelaying the onset or development or progression of the disease ordisorder.

As used herein, a subject is “in need of” a treatment if such subjectwould benefit biologically, medically or in quality of life from suchtreatment.

As used herein, the term “dyslipidemia” refers to an abnormality in, orabnormal amounts of lipids and lipoproteins in the blood and the diseasestates resulting, caused by, exacerbated by, or adjunct to suchabnormality (see, Dorland's Illustrated Medical Dictionary, 29th editionor subsequent versions thereof, W.B. Saunders Publishing Company, NewYork, N.Y.). Disease states encompassed within the definition ofdyslipidemia as used herein include hyperlipidemia,hypertriglyceridemia, low plasma HDL, high plasma LDL, high plasma VLDL,liver cholestasis, and hypercholesterolemia.

As used herein, the phrase “diseases related to dyslipidemia” as usedherein refers to diseases including but not limited to atherosclerosis,thrombosis, coronary artery disease, stroke, and hypertension. Diseasesrelated to dyslipidemia also include metabolic diseases such as obesity,diabetes, insulin resistance, and complications thereof.

As used herein, the term “cholestasis” refers to any condition in whichthe flow of bile from the liver is blocked, and may be intrahepatic(i.e., occurring inside the liver) or extrahepatic (i.e., occurringoutside the liver).

As used herein, “liver fibrosis” includes liver fibrosis due to anycause, including but not limited to virally-induced liver fibrosis suchas that due to hepatitis B and C; exposure to alcohol (alcoholic liverdisease), pharmaceutical compounds, oxidative stress, cancer radiationtherapy or industrial chemicals; and diseases such as primary biliarycirrhosis, fatty liver, obesity, non-alcoholic steatohepatitis, cysticfibrosis, hemochromatosis, and auto-immune hepatitis.

As used herein, “FXR agonist” refers to an agent that directly binds toand upregulates the activity of FXR.

As used herein, the term “a,” “an,” “the” and similar terms used in thecontext of the present invention (especially in the context of theclaims) are to be construed to cover both the singular and plural unlessotherwise indicated herein or clearly contradicted by the context.

Any definition herein may be used in combination with any otherdefinition to describe a composite structural group. By convention, thetrailing element of any such definition is that which attaches to theparent moiety. For example, the composite group alkoxyalkyl wouldrepresent an alkoxy group attached to the parent molecule through analkyl group.

MODES OF CARRYING OUT THE INVENTION

The present invention relates to compositions and methods for FXR.Various embodiments of the invention are described herein. It will berecognized that features specified in each embodiment may be combinedwith other specified features to provide further embodiments.

In a first embodiment, the compounds of the present invention aredefined by Formula (I):

or a pharmaceutical acceptable salt thereof, wherein,

R⁰ is Ring A or C₁₋₆ alkyl;

Ring A is aryl; 5-10 membered heteroaryl comprising 1-3 N, O or Sheteroatoms; 4-6 membered heterocycle comprising 1-2 N, O or Sheteroatoms; or C₃₋₇ cycloalkyl; and said Ring A is unsubstituted orsubstituted by 1-2 substituents independently represented by R²;

Ring B is aryl; 5-10 membered heteroaryl comprising 1-3 N, O or Sheteroatoms; 4-6 membered heterocycle comprising 1-2 N, O or Sheteroatoms; or C₃₋₇ cycloalkyl; and said Ring B is unsubstituted orsubstituted by 1-2 substituents independently represented by R²;

X is —(CR⁴R⁵)— or —C(O)—;

Y is —O—, —(CR⁴R⁵)—, *—O(CR⁴R⁵)— or —NR—, wherein “*” indicates thepoint of attachment of Y to the ring containing the Z ring atoms;

Z¹, Z², Z³, and Z⁴ are each independently —CR³— or —N—;

L¹ is *¹—(CR⁴R⁵)₁₋₂— or *¹—(CR⁴R⁵)—C(O)—NR—, wherein “*¹” indicates thepoint of attachment of L¹ to N;

L² is *²—(CR⁴R⁵)₁₋₂—, *²—(CR⁴R⁵)—C(O)—, *²—(CR⁴R⁵)—C(O)—NR—,*²—(CR⁴R⁵)₂—O—, *²—(CR⁴R⁵)₂—NR—, *²—(CR⁴R⁵)₂—SO₂—, *²—(CR⁴R⁵)₂—NR—C(O)—,or *²—(CR⁴R⁵)—C(O)—NR—(CR⁴R⁵)—; wherein “*²” indicates the point ofattachment of L² to N;

L³ is —(CR⁴R⁵)— or —C(O)—;

each R² is independently halo, hydroxyl, C₁₋₆ alkyl, or halo-substitutedC₁₋₆ alkyl;

each R³ is independently hydrogen, halo, or C₁₋₆ alkyl; and

R, R⁴ and R⁵ are independently hydrogen or C₁₋₆ alkyl.

In a second embodiment, the compounds of the present invention aredefined by Formula (I) as defined in the first embodiment, wherein

-   -   R⁰ is Ring A or C₁₋₆ alkyl;    -   Ring A is aryl; 5-10 membered heteroaryl comprising 1-3 N, O or        S heteroatoms; or C₃₋₇ cycloalkyl; and said Ring A is        unsubstituted or substituted by 1-2 substituents independently        represented by R²;    -   Ring B is aryl; 5-10 membered heteroaryl comprising 1-3 N, O or        S heteroatoms; or C₃₋₇ cycloalkyl; and said Ring B is        unsubstituted or substituted by 1-2 substituents independently        represented by R²;    -   X is —(CR⁴R⁵)—;    -   Y is —O—, —(CR⁴R⁵)—, or *—O(CR⁴R⁵)—, wherein “*” indicates the        point of attachment of Y to the ring containing the Z ring        atoms;    -   Z¹, Z², Z³, and Z⁴ are each independently —CR³— or —N—;    -   L¹ is *¹—(CR⁴R⁵)₁₋₂— wherein “*¹” indicates the point of        attachment of L¹ to N;    -   L² is *²—(CR⁴R⁵)₁₋₂—, *²—(CR⁴R⁵)—C(O)—NR—, *²—(CR⁴R⁵)₂—O—,        *²—(CR⁴R⁵)₂—NR— or *²—(CR⁴R⁵)—C(O)—NR—(CR⁴R⁵)—; wherein “*²”        indicates the point of attachment of L² to N;    -   L³ is —C(O)—;    -   each R² is independently halo, C₁₋₆ alkyl, or halo-substituted        C₁₋₆ alkyl;    -   each R³ is independently hydrogen, halo, or C₁₋₆ alkyl;    -   R, R⁴ and R⁵ are independently hydrogen or C₁₋₆ alkyl.

In a third embodiment, the compounds of the present invention aredefined by Formula (II):

or a pharmaceutically acceptable salt thereof; wherein,

-   -   R⁰ is Ring A or C₁₋₆ alkyl; wherein Ring A is phenyl, pyridyl or        cyclopropyl, each of which is unsubstituted or substituted by        1-2 substituents independently represented by R²;    -   Ring B is selected from phenyl, pyridyl, 1H-indolyl, and C₃₋₇        cycloalkyl, each of which is unsubstituted or substituted by 1-2        substituents independently represented by R²;    -   X is —(CR⁴R⁵)—;    -   Y is —O—, —(CR⁴R⁵)— or *—O(CR⁴R⁵)—, wherein “*” indicates the        point of attachment of Y to the ring containing the Z ring        atoms;    -   Z¹, Z², Z³, and Z⁴ are each independently —CR³— or —N—;    -   L¹ is *¹—(CR⁴R⁵)₁₋₂— wherein “*¹” indicates the point of        attachment of L¹ to N;    -   L² is *²—(CR⁴R⁵)₁₋₂—, *²—(CR⁴R⁵)—C(O)—NR—, *²—(CR⁴R⁵)₂—O—,        *²—(CR⁴R⁵)₂—NR— or *²—(CR⁴R⁵)—C(O)—NR—(CR⁴R⁵)—; where “*²”        indicates the point of attachment of L² to N;    -   each R² is independently halo, C₁₋₆ alkyl, or halo-substituted        C₁₋₆ alkyl;    -   each R³ is independently hydrogen, halo, or C₁₋₆ alkyl; and    -   R, R⁴ and R⁵ are independently hydrogen or C₁₋₆ alkyl.

In a fourth embodiment, R⁰ in any of the above embodiments is selectedfrom *³—CH₂C(CH₃)₂—, *³—CH₂CH(CH₃)—, and *³-cyclopropane-1,1,-diyl,wherein “*³” indicates the point of attachment of R⁰ to L².

With reference to any one of the above embodiments, Y can be selectedfrom —O—, —CH₂—, —C(CH₃)₂—, and *—O—CH₂—, wherein “*” indicates thepoint of attachment of Y to the six membered ring containing the Z ringatoms.

With reference to any one of the above embodiments, L¹ can be —(CR⁴R⁵)—or —CH₂—. In one variation, L¹ is —(CR⁴R⁵)—.

With reference to any one of the above embodiments, L² can be selectedfrom —(CH₂)—, *²—CH₂C(O)NH—, *²—CH(CH₃)C(O)NH—, *²—CH₂C(O)NHCH₂—,*²—(CH₂)₂O—, and *²—(CH₂)₂NH—, wherein “*²” indicates the point ofattachment of L² to N.

With reference to any one of the above embodiments, each R² when presentcan independently be selected from halo, methyl, and trifluoromethyl.

In a fifth embodiment, the compounds of the present invention arerepresented by Formula (III):

or a pharmaceutically acceptable salt thereof; wherein

-   -   Ring A is phenyl or pyridyl, each of which is unsubstituted or        substituted by 1-2 substituents independently represented by R²;    -   Ring B is selected from phenyl, pyridyl, 1H-indolyl, and        cyclopentyl, each of which is unsubstituted or substituted by        1-2 substituents independently represented by R²;    -   L¹ is —(CR⁴R⁵)—,    -   L² is selected from —(CH₂)—, *²—CH₂C(O)NH—, *²—CH(CH₃)C(O)NH—,        *²—CH₂C(O)NHCH₂—, *²—(CH₂)₂O—, and *²—(CH₂)₂NH—; wherein “*²”        indicates the point of attachment of L² to N;    -   X is CH₂;    -   Y is selected from —O—, —CH₂—, —C(CH₃)₂— and *—O—CH₂—, wherein        “*” indicates the point of attachment of Y to the ring        containing the Z ring atoms;    -   Z¹ is CR³ or N;    -   Z² is CR³;    -   Z³ is CR³;    -   Z⁴ is CR³ or N;    -   each R² is independently selected from halo, methyl, and        trifluoromethyl;    -   each R³ is independently hydrogen, halo, or C₁₋₆ alkyl; and    -   each of R⁴ and R⁵ is independently hydrogen or methyl.

In a sixth embodiment, Y in any of the above embodiments is —O—. In analternative embodiment, Y is —CH₂—. In another alternative embodiment, Yis *—O—CH₂—, wherein “*” indicates the point of attachment of Y to thering containing the Z ring atoms.

In a seventh embodiment, L¹ in any of the above embodiments is —CH₂—.

In an eighth embodiment, L² in any of the above embodiments is selectedfrom —(CH₂)—, *²—CH₂C(O)NH—, *²—(CH₂)₂O—, and *²—(CH₂)₂NH—; wherein “*²”indicates the point of attachment of L² to N.

In a ninth embodiment, L² in any of the above embodiments is —(CH₂)—.

In a tenth embodiment, R² when present in any one of the aboveembodiments is independently fluoro or methyl. In one variation, each R²is fluoro. In another variation, each R² is methyl.

In an eleventh embodiment, R³ in any of the above embodiments isselected from hydrogen, fluoro, chloro, and methyl.

In a twelfth embodiment, with reference to any one of the aboveembodiments, Z¹ is selected from CH, CF, CCH₃, and N; Z² is selectedfrom CH, CF, CCl, and CCH₃; Z³ is selected from CH, CF, CCl, and CCH₃;and Z⁴ is CH or N.

With reference to any one of the above embodiments, each of R, R⁴, R⁵and R⁶ when present, can independently be hydrogen or methyl. In onevariation, R, R⁴, R⁵ and R⁶ when present, is hydrogen.

In a thirteenth embodiment, the compounds of the present invention arerepresented by Formula (IV):

-   -   each phenyl ring is optionally further substituted by 1-2        substituents independently represented by R², wherein R² is        fluoro or methyl;    -   L² is selected from —CH₂—, *²—CH₂CH₂NH—, *²—CH₂CH₂O—, and        *²—CH₂C(O)NH—, wherein “*²” indicates the point of attachment of        L² to N;    -   Z² is selected from CH, CF, CCl, and CCH₃; and    -   Z³ is selected from CH, CF, CCl, and CCH₃.

In one embodiment, L² in any of the above embodiments is —(CH₂)—. In analternative embodiment, L² is *²—(CH₂)₂NH—. In another alternativeembodiment, L² is *²—CH₂C(O)NH—. In each embodiment, “*²” indicates thepoint of attachment of L² to N.

Particular compounds according to the present invention include, but arenot limited to:4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(8-chloro-1-methyl-N-(3-methylbenzyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;4-fluoro-3-(2-(1-methyl-N-(3-methylbenzyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(N-(2-fluorobenzyl)-1,8-dimethyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid; 3-(2-(N-(2-fluorobenzyl)-1,6-dimethyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(7-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(7-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(6,8-difluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1,7-dimethyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1,6-dimethyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(8-chloro-6-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;4-fluoro-3-(2-(N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(7,8-difluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-(2-(7,8-difluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-(2-(N-benzyl-7,8-difluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;4-fluoro-3-(2-(8-fluoro-1-methyl-N-(3-methylbenzyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;4-fluoro-3-(2-(N-(2-fluorobenzyl)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinoline-3-carboxamido)acetamido)benzoicacid; 4-fluoro-3-(2-(N-(2-fluorobenzyl)-1-methyl-4,5-dihydro-1H-pyrazolo[3,4-f]quinoline-3-carboxamido)acetamido)benzoic acid;(S)-4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)propanamido)benzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-4,5-dihydro-1H-benzo[g]indazole-3-carboxamido)acetamido)benzoicacid;3-(2-(8-chloro-7-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;4-fluoro-3-(2-(N-(3-fluorobenzyl)-1,5,5-trimethyl-4,5-dihydro-1H-benzo[g]indazole-3-carboxamido)acetamido)benzoicacid;4-fluoro-3-(2-(N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-(2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-(2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-(2-(N-benzyl-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-(2-(9-chloro-N-(2-fluorobenzyl)-1-methyl-4,5-dihydro-1H-benzo[2,3]oxepino[4,5-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(8-chloro-N-(cyclopentylmethyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-(2-(8-chloro-N-(cyclopentylmethyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-5-fluorobenzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-methylbenzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-2-methylpropanoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-2,2-dimethylpropanoicacid;1-((2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)methyl)cyclopropanecarboxylic acid;4-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)benzoicacid;4-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)benzoicacid;N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-methylbenzoicacid;4-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3,5-dimethylbenzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-4-fluorobenzoicacid;4-(2-(8-chloro-N-(cyclopentylmethyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3,5-dimethylbenzoicacid;4-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3,5-difluorobenzoicacid;4-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-(trifluoromethyl)benzoicacid;4-(2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3,5-difluorobenzoicacid;3,5-difluoro-4-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)benzoicacid;4-(2-(N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3,5-difluorobenzoicacid;4-(2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid;4-(2-(N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid;4-(2-(N-benzyl-7,8-difluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid;3-fluoro-4-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)benzoicacid;4-(2-(7,8-difluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid;4-(2-(8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid;3-fluoro-4-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)benzoicacid;4-(2-(7,8-difluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid;3-((2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-((2-(8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid;3-((2-(8-chloro-1-methyl-N-(3-methylbenzyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid;4-fluoro-3-((2-(8-fluoro-1-methyl-N-(3-methylbenzyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid;3-((2-(7,8-difluoro-1-methyl-N-(3-methylbenzyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid;4-fluoro-3-((2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid;4-fluoro-3-((2-(N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid;3-((2-(N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid;3-((2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid;3-((2-(7,8-difluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid;3-((2-(N-benzyl-7,8-difluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid;4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)-3-fluorobenzoicacid;4-((8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-N-(3,5-difluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)-3-fluorobenzoicacid;4-((8-chloro-1-methyl-N-(3-methylbenzyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-N-(2,3-difluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-N-(3-fluoro-5-methylbenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((N-(3,5-difluorobenzyl)-8-fluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-1-methyl-N-(3-methylbenzyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)-3-fluorobenzoicacid;3-fluoro-4-((8-fluoro-1-methyl-N-(3-methylbenzyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((N-((1H-indol-5-yl)methyl)-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;5-((8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)picolinicacid;4-((8-chloro-N-((5-fluoropyridin-3-yl)methyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-N-((5-chloropyridin-3-yl)methyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)-2-fluorobenzoicacid;4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)-2-fluorobenzoicacid;N-benzyl-N-(4-carbamoylbenzyl)-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamide;4-((8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)-3-fluorobenzoicacid;3-fluoro-4-((8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)-3-fluorobenzoicacid;4-((8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)-2-fluorobenzoicacid;6-((8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)nicotinicacid;5-((8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)-6-methylpicolinicacid;4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid; and3-((2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid; or a pharmaceutically acceptable salt, tautomer or stereoisomerthereof.

More particular examples of the compounds according to the presentinvention include but are not limited to:4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(7-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid;3-(2-(7,8-difluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid;3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)benzoicacid;3-((2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid;3-((2-(8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid;4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid;4-((8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid; and4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid; or a pharmaceutically acceptable salt, tautomer or stereoisomerthereof.

In one embodiment, the present invention provides a compound accordingto any one of the above embodiments, wherein the compound is in the formof a pharmaceutically acceptable salt selected from TRIS(2-amino-2-hydroxymethyl-1,3-propanediol), arginine, lysine, sodium andmeglumine salt.

The present invention also provides a compound of Formula (V):

-   -   wherein each phenyl ring is unsubstituted or substituted by 1-2        substituents independently represented by R², wherein R² is        fluoro or methyl;    -   L² is selected from *²—CH₂—, *²—CH₂CH₂NH—, *²—CH₂CH₂O—, and        *²—CH₂C(O)NH—, wherein “*²” indicates the point of attachment of        L² to N;    -   Z² is selected from CH, CF, CCl, and CCH₃;    -   Z³ is selected from CH, CF, CCl, and CCH₃; and    -   R⁶ is C₁₋₆ alkyl.

In another aspect, the present invention provides a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundaccording to any one of the above embodiments and variations or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier, diluent or excipient.

In yet another aspect, the present invention provides a combinationcomprising a therapeutically effective amount of a compound according toany one of the above embodiments and variations or a pharmaceuticallyacceptable salt thereof, and a second therapeutic agent.

In still another aspect, the invention provides a method for treating acondition mediated by farnesoid X receptors (FXR) in a subject sufferingtherefrom, comprising administering to the subject a therapeuticallyeffective amount of a compound of any one of the above embodiments andvariations or a pharmaceutically acceptable salt thereof; and optionallyin combination with a second therapeutic agent.

In still another aspect, the invention provides a compound according toany of the above embodiments and variations or a pharmaceuticallyacceptable salt thereof, and optionally in combination with a secondtherapeutic agent, for use in treating a condition mediated by FXR.

In yet another aspect, the invention provides the use of a compound ofany one of the above embodiments and variations or a pharmaceuticallyacceptable salt thereof, and optionally in combination with a secondtherapeutic agent, for the preparation of a medicament for the treatmentof a condition mediated by FXR in a subject.

In one embodiment, the condition mediated by FXR with respect to any ofthe above methods, uses or combinations, is a liver disease or agastrointestinal disease. For example, the compounds of the inventionmay be used for treating a liver disease mediated by FXR, wherein theliver disease is selected from cholestasis (e.g., intrahepaticcholestasis, estrogen-induced cholestasis, drug-induced cholestasis,cholestasis of pregnancy, parenteral nutrition-associated cholestasis,progressive familiar cholestasis (PFIC)); Alagille syndrome, primarybiliary cirrhosis (PBC), primary sclerosing cholangitis, ductopenicliver transplant rejection, liver transplant associated graft versushost disease, cystic fibrosis liver disease, non-alcoholic fatty liverdisease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic liverdisease, and parenteral nutrition-associated liver disease. Thecompounds of the invention may be also be used for treating agastrointestinal disease mediated by FXR, wherein the gastrointestinaldisease is selected from bile acid malabsorption (including primary bileacid diarrhea and secondary bile acid diarrhea), bile reflux gastritis,and inflammatory bowel disease such as Crohn's disease, ulcerativecolitis, collagenous colitis, lymphocytic colitis, diversion colitis,indeterminate colitis and Behçet's disease.

More particularly, the condition mediated by FXR is non-alcoholic fattyliver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). Withreference to combination therapies of the invention, the othertherapeutic agent can also be useful in the treatment of non-alcoholicfatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).

In one embodiment, the compounds of the invention are administeredenterally; and more particularly, orally.

Unless specified otherwise, the term “compounds of the presentinvention” refers to compounds of Formula I, II, III, IV or V,pharmaceutically acceptable salt thereof, prodrugs, and inherentlyformed moieties (e.g., polymorphs, solvates and/or hydrates). Thecompounds of the present invention may be stereoisomers (includingdiastereoisomers and enantiomers), and may be a mixture of stereoisomersor a single stereoisomer. The compounds of the present invention mayalso be tautomers and isotopically labeled compounds (includingdeuterium substitutions). Further compounds of the invention aredetailed in the Examples, infra.

Certain of the compounds described herein contain one or more asymmetriccenters or axes and may thus give rise to enantiomers, diastereomers,and other stereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-. The present invention is meant toinclude all possible isomers, including racemic mixtures, optically pureforms and intermediate mixtures. Optically active (R)- and (S)-isomersmay be prepared using chiral synthons or chiral reagents, or resolvedusing conventional techniques. If the compound contains a double bond,the substituent may be E or Z configuration. If the compound contains adisubstituted cycloalkyl, the cycloalkyl substituent may have a cis- ortrans-configuration. All tautomeric forms are also intended to beincluded.

Any formula given herein is also intended to represent unlabeled formsas well as isotopically labeled forms of the compounds. Isotopicallylabeled compounds have structures depicted by the formulas given hereinexcept that one or more atoms are replaced by an atom having a selectedatomic mass or mass number. Examples of isotopes that can beincorporated into compounds of the invention include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine,such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸F, ³¹P, ³²P, ³⁵S, ³⁶Cl and ¹²⁵Irespectively. The invention includes various isotopically labeledcompounds as defined herein, for example those into which radioactiveisotopes, such as ³H, ¹³C, and ¹⁴C, are present. Such isotopicallylabelled compounds are useful in metabolic studies (with ¹⁴C), reactionkinetic studies (with, for example ²H or ³H), detection or imagingtechniques, such as positron emission tomography (PET) or single-photonemission computed tomography (SPECT) including drug or substrate tissuedistribution assays, or in radioactive treatment of patients. Inparticular, an ¹⁸F or labeled compound may be particularly desirable forPET or SPECT studies. Isotopically labeled compounds of this inventionand prodrugs thereof can generally be prepared by carrying out theprocedures disclosed in the schemes or in the examples and preparationsdescribed below by substituting a readily available isotopically labeledreagent for a non-isotopically labeled reagent.

Further, substitution with heavier isotopes, particularly deuterium(i.e., ²H or D) may afford certain therapeutic advantages resulting fromgreater metabolic stability, for example increased in vivo half-life orreduced dosage requirements or an improvement in therapeutic index. Itis understood that deuterium in this context is regarded as asubstituent of a compound of the Formula (I). The concentration of sucha heavier isotope, specifically deuterium, may be defined by theisotopic enrichment factor. The term “isotopic enrichment factor” asused herein means the ratio between the isotopic abundance and thenatural abundance of a specified isotope. If a substituent in a compoundof this invention is denoted deuterium, such compound has an isotopicenrichment factor for each designated deuterium atom of at least 3500(52.5% deuterium incorporation at each designated deuterium atom), atleast 4000 (60% deuterium incorporation), at least 4500 (67.5% deuteriumincorporation), at least 5000 (75% deuterium incorporation), at least5500 (82.5% deuterium incorporation), at least 6000 (90% deuteriumincorporation), at least 6333.3 (95% deuterium incorporation), at least6466.7 (97% deuterium incorporation), at least 6600 (99% deuteriumincorporation), or at least 6633.3 (99.5% deuterium incorporation).

Isotopically-labeled compounds of Formula I, II, III, IV and V cangenerally be prepared by conventional techniques known to those skilledin the art or by processes analogous to those described in theaccompanying Examples and Processes using an appropriateisotopically-labeled reagent in place of the non-labeled reagentpreviously employed.

Pharmaceutically acceptable solvates in accordance with the inventioninclude those wherein the solvent of crystallization may be isotopicallysubstituted, e.g. D₂O, d₆-acetone, d₆-DMSO.

Compounds of the invention, i.e. compounds of Formula I, II, III, IV andV that contain groups capable of acting as donors and/or acceptors forhydrogen bonds may be capable of forming co-crystals with suitableco-crystal formers. These co-crystals may be prepared from compounds ofFormula I, II, III, IV or V by known co-crystal forming procedures. Suchprocedures include grinding, heating, co-subliming, co-melting, orcontacting in solution compounds of Formula I, II, III, IV or V with theco-crystal former under crystallization conditions and isolatingco-crystals thereby formed. Suitable co-crystal formers include thosedescribed in WO 2004/078163. Hence the invention further providesco-crystals comprising a compound of Formula I, II, III, IV or V.

Any asymmetric atom (e.g., carbon or the like) of the compound(s) of thepresent invention can be present in racemic or enantiomericallyenriched, for example the (R)-, (S)- or (R,S)-configuration. In certainembodiments, each asymmetric atom has at least 50% enantiomeric excess,at least 60% enantiomeric excess, at least 70% enantiomeric excess, atleast 80% enantiomeric excess, at least 90% enantiomeric excess, atleast 95% enantiomeric excess, or at least 99% enantiomeric excess inthe (R)- or (S)-configuration. Substituents at atoms with unsaturatedbonds may, if possible, be present in cis-(Z)- or trans-(E)-form.

Accordingly, as used herein a compound of the present invention can bein the form of one of the possible isomers, rotamers, atropisomers,tautomers or mixtures thereof, for example, as substantially puregeometric (cis or trans) isomers, diastereomers, optical isomers(antipodes), racemates or mixtures thereof. Any resulting mixtures ofisomers can be separated on the basis of the physicochemical differencesof the constituents, into the pure or substantially pure geometric oroptical isomers, diastereomers, racemates, for example, bychromatography and/or fractional crystallization. Any resultingracemates of final products or intermediates can be resolved into theoptical antipodes by known methods, e.g., by separation of thediastereomeric salts thereof, obtained with an optically active acid orbase, and liberating the optically active acidic or basic compound. Inparticular, a basic moiety may thus be employed to resolve the compoundsof the present invention into their optical antipodes, e.g., byfractional crystallization of a salt formed with an optically activeacid, e.g., tartaric acid, dibenzoyl tartaric acid, diacetyl tartaricacid, di-O,O′-p-toluoyl tartaric acid, mandelic acid, malic acid orcamphor-10-sulfonic acid. Racemic products can also be resolved bychiral chromatography, e.g., high pressure liquid chromatography (HPLC)using a chiral adsorbent.

Pharmacology and Utility

The compounds of Formula I, II, III, IV and V in free form or in saltform, exhibit valuable pharmacological properties, e.g. FXR modulatingproperties, e.g. as indicated in in vitro and/or in vivo tests asprovided in the next sections, and are therefore indicated for therapyin treating a disorder which may be treated by modulating FXR, such asthose described below.

With the development of the first synthetic FXR ligand GW4064 as a toolcompound (Maloney et al., J. Med. Chem. 2000, 43(16), 2971-2974; Willsonet al., Med. Res. Rev. 2001, 21(6) 513-22), and the development of thesemisynthetic artificial bile acid ligand 6-alpha-ethyl-CDCA, theeffects of superstimulation of FXR by potent agonists could be analyzed.It was shown that both ligands induce bile flow in bile duct ligatedanimals. In addition to choleretic effects, hepatoprotective effectscould also be demonstrated (Pellicciari et al., J. Med. Chem. 2002,45(17), 3569-3572; Liu et al., J. Clin. Invest. 2003, 112(11),1678-1687). This hepatoprotective effect was further narrowed down to ananti-fibrotic effect that results from the repression of TissueInhibitors of Matrix-Metalloproteinases, TIMP-1 and 2, the induction ofcollagen-deposit resolving Matrix-Metalloproteinase 2 (MMP-2) in hepaticstellate cells and the subsequent reduction of alpha-collagen mRNA andTransforming growth factor beta (TGF-beta) mRNA which are bothpro-fibrotic factors by FXR agonists (Fiorucci et al., Gastroenterology2004, 127(5), 1497-1512; Fiorucci et al., Pharmacol. Exp. Ther. 2005,314(2), 584-595).

The anti-fibrotic activity of FXR is at least partially mediated by theinduction of PPARγ, a further nuclear receptor, with which anti-fibroticactivity is associated (Fiorucci et al., J. Pharmacol. Exp. Ther. 2005,315(1), 58-68; Galli et al., Gastroenterology 2002, 122(7), 1924-1940;Pineda Torra et al., Mol. Endocrinol. 2003, 17(2), 259-272).Furthermore, anti-cholestatic activity was demonstrated in bile-ductligated animal models as well as in animal models of estrogen-inducedcholestasis (Fiorucci et al., J. Pharmacol. Exp. Ther. 2005, 313(2),604-612).

Genetic studies demonstrate that in hereditary forms of cholestasis(Progressive Familiar Intrahepatic Cholestasis=PFIC, Type I-IV), eithernuclear localization of FXR itself is reduced as a consequence of amutation in the FIC1 gene (in PFIC Type I, also called Byler's Disease)(Chen et al., Gastroenterology. 2004, 126(3), 756-64; Alvarez et al.,Hum. Mol. Genet. 2004; 13(20), 2451-60) or levels of the FXR target geneencoding MDR-3 phospholipid export pump are reduced (in PFIC Type III).Taken together, there is a growing body of evidence that FXR bindingcompounds will demonstrate substantial clinical utility in thetherapeutic regimen of chronic cholestatic conditions such as PrimaryBiliary Cirrhosis (PBC) or Primary Sclerosing Cholangitis (PSC)(reviewed in: Rizzo et al., Curr. Drug Targets Immune Endocr. Metabol.Disord. 2005, 5(3), 289-303; Zollner, Mol. Pharm. 2006, 3(3), 231-51,Cai et al., Expert Opin. Ther. Targets 2006, 10(3), 409-421).

Furthermore, FXR seems to be involved in the regulation of many diversephysiological processes which are relevant in the etiology and for thetreatment of diseases as diverse as cholesterol gallstones, metabolicdisorders such as Type II Diabetes, dyslipidemias or obesity, chronicinflammatory diseases such as Inflammatory Bowel Diseases or chronicintrahepatic forms of cholestasis and many others diseases (Claudel etal., Arterioscler. Thromb. Vase. Biol. 2005, 25(10), 2020-2030; Westinet al., Mini Rev. Med. Chem. 2005, 5(8), 719-727).

FXR has also been shown to be a key regulator of serum triglycerides(Maloney et al., J. Med. Chem. 2000, 43(16), 2971-2974; Willson et al.,Med. Res. Rev. 2001, 21(6), 513-22). Recent reports indicate thatactivation of FXR by synthetic agonists leads to significant reductionof serum triglycerides, mainly in the form of reduced VLDL, but also toreduced total serum cholesterol (Figge et al., J. Biol. Chem. 2004,279(4), 2790-2799; Bilz et al., Am. J. Physiol. Endocrinol. Metab. 2006,290(4), E716-22). However, the lowering of serum triglycerides is not astand-alone effect. Treatment of db/db or ob/ob mice with synthetic FXRagonist GW4064 resulted in marked and combined reduction of serumtriglycerides, total cholesterol, free fatty acids, and ketone bodiessuch as 3-OH Butyrate. Moreover, FXR activation engages with theintracellular insulin signaling pathway in hepatocytes, resulting inreduced output of glucose from liver gluconeogenesis but concomitantincrease in liver glycogen. Insulin sensitivity as well as glucosetolerance were positively impacted by FXR treatment (Stayrook et al.,Endocrinology 2005, 146(3), 984-91; Zhang et al., Proc. Natl. Acad. Sci.USA 2006, 103(4), 1006-1011; Cariou et al., J. Biol. Chem. 2006, 281,11039-11049; Ma et al., J. Clin. Invest. 2006, 116(4), 1102-1109;Duran-Sandoval et al., Biochimie 2005, 87(1), 93-98).

The compounds of the invention are also useful for the treatment ofgastrointestinal diseases, including but not limited to bile acidmalabsorption (including primary bile acid diarrhea and secondary bileacid diarrhea), bile reflus gastritis and inflammatory bowel diseases(IBD). Bile acid malabsorption, which leads to excessive fecal bile acidexcretion and diarrhea in patients, is characterized by a cycle whereinthe feedback regulation of bile acid synthesis is interrupted, resultingin additional bile acid production. Feedback regulation of bile acidsynthesis is under the control of an endocrine pathway, whereinactivation of the nuclear bile acid receptor FXR induces entericexpression of fibroblast growth factor 15 (FGF15) in rodents or FGF19 inhumans. In liver, FGF15 or FGF19 act together with FXR-mediatedexpression of small heterodimer partner to repress bile acid synthesis(Jung et al., Journal of Lipid Research 48: 2693-2700 (2007) Walters JR, Nat Rev Gastroenterol Hepatol. 11(7):426-34 (2014)).

In another embodiment, the compounds according to the invention areuseful for beneficially altering lipid profiles, including but notlimited to lowering total cholesterol levels, lowering LDL cholesterollevels, lowering VLDL cholesterol levels, raising HDL cholesterollevels, and/or lowering triglyceride levels. Thus, the present inventionprovides a method for treating FXR mediated conditions such asdyslipidemia and diseases related to dyslipidemia comprisingadministering a therapeutically effective amount of a compound of thepresent invention to a subject in need thereof.

In a further embodiment, the compound or pharmaceutical composition isused for treating a disease selected from the group consisting of lipidand lipoprotein disorders such as hypercholesterolemia,hypertriglyceridemia, and atherosclerosis as a clinically manifestcondition which can be ameliorated by FXR's beneficial effect on raisingHDL cholesterol, lowering serum triglycerides, increasing conversion ofliver cholesterol into bile acids and increased clearance and metabolicconversion of VLDL and other lipoproteins in the liver.

In one further embodiment, said compound and pharmaceutical compositionare used for the preparation of a medicament where the combined lipidlowering, anti-cholestatic and anti-fibrotic effects of FXR-targetedmedicaments can be exploited for the treatment of liver steatosis andassociated syndromes such as non-alcoholic steatohepatitis (NASH), orfor the treatment of cholestatic and fibrotic effects that areassociated with alcohol-induced cirrhosis, or with viral-borne forms ofhepatitis.

FXR seems also to be involved in the control of antibacterial defense inthe intestine (Inagaki et al., Proc. Natl. Acad. Sci. USA. 2006,103(10), 3920-3905), and may have a beneficial impact in the therapy ofInflammatory Bowel Disorders (IBD), particularly those forms where theupper (ileal) part of the intestine is affected (e.g. ileal Crohn'sdisease) because this seems to be the site of action of FXR's control onbacterial growth. In IBD, the desensitization of the adaptive immuneresponse is somehow impaired in the intestinal immune system. Bacterialovergrowth might then be the causative trigger towards establishment ofa chronic inflammatory response. Hence, dampening of bacterial growth byFXR-borne mechanisms might be a key mechanism to prevent acuteinflammatory episodes. Thus, the invention also relates to a compoundaccording to formula (I) or a pharmaceutical composition comprising saidcompound for treating a disease related to Inflammatory Bowel Diseasessuch as Crohn's disease or ulcerative colitis. FXR-mediated restorationof intestinal barrier function and reduction in non-commensal bacterialload is believed to be helpful in reducing the exposure of bacterialantigens to the intestinal immune system and can therefore reduceinflammatory responses.

The invention further relates to a compound or pharmaceuticalcomposition for the treatment of obesity and associated disorders suchas metabolic syndrome (combined conditions of dyslipidemias, diabetesand abnormally high body-mass index), which can be overcome byFXR-mediated lowering of serum triglycerides, blood glucose andincreased insulin sensitivity and FXR-mediated weight loss. Thecompounds or pharmaceutical composition of the present invention arealso useful in the preparation of a medicament for treating clinicalcomplications of Type I and Type II Diabetes such as diabeticnephropathy, diabetic retinopathy, and Peripheral Arterial OcclusiveDisease (PAOD).

Furthermore, conditions and diseases which result from chronic fatty andfibrotic degeneration of organs due to enforced lipid and specificallytriglyceride accumulation and subsequent activation of profibroticpathways may also be treated by applying the compounds or pharmaceuticalcomposition of the present invention. Such conditions and diseasesencompass Non-Alcoholic Steatohepatitis (NASH) and chronic cholestaticconditions in the liver, glomerulosclerosis and diabetic nephropathy inthe kidney, macular degeneration and diabetic retinopathy in the eye andneurodegenerative diseases such as Alzheimer's disease in the brain ordiabetic neuropathies in the peripheral nervous system.

Examples of other FXR-mediated disease include drug-induced bile ductinjury, bile duct obstruction, gallstones, cholelithiasis, liverfibrosis, liver cirrhosis, alcohol-induced cirrhosis, dyslipidemia,atherosclerosis, diabetes, diabetic nephropathy, colitis, newbornjaundice, prevention of kernicterus, veno-occlusive disease, portalhypertension, metabolic syndrome, hypercholesterolemia, intestinalbacterial overgrowth, erectile dysfunction, and other FXR-mediatedconditions leading to extrahepatic cholestasis.

Administration and Pharmaceutical Compositions

In another aspect, the present invention provides a pharmaceuticalcomposition comprising a compound of the present invention and apharmaceutically acceptable carrier. The pharmaceutical composition canbe formulated for particular routes of administration such as oraladministration, parenteral administration, and rectal administration,etc. In addition, the pharmaceutical compositions of the presentinvention can be made up in a solid form (including without limitationcapsules, tablets, pills, granules, powders or suppositories), or in aliquid form (including without limitation solutions, suspensions oremulsions). The pharmaceutical compositions can be subjected toconventional pharmaceutical operations such as sterilization and/or cancontain conventional inert diluents, lubricating agents, or bufferingagents, as well as adjuvants, such as preservatives, stabilizers,wetting agents, emulsifiers and buffers, etc.

In a particular embodiment, the pharmaceutical composition is formulatedfor oral administration. Typically, the pharmaceutical compositions aretablets or gelatin capsules comprising the active ingredient togetherwith

a) diluents, e.g., lactose, dextrose, sucrose, mannitol, sorbitol,cellulose and/or glycine;

b) lubricants, e.g., silica, talcum, stearic acid, its magnesium orcalcium salt and/or polyethyleneglycol; for tablets also

c) binders, e.g., magnesium aluminum silicate, starch paste, gelatin,tragacanth, methylcellulose, sodium carboxymethylcellulose and/orpolyvinylpyrrolidone; if desired

d) disintegrants, e.g., starches, agar, alginic acid or its sodium salt,or effervescent mixtures; and/or

e) absorbents, colorants, flavors and sweeteners.

Tablets may be either film coated or enteric coated according to methodsknown in the art.

Suitable compositions for oral administration include an effectiveamount of a compound of the invention in the form of tablets, lozenges,aqueous or oily suspensions, dispersible powders or granules, emulsion,hard or soft capsules, or syrups or elixirs. Compositions intended fororal use are prepared according to any method known in the art for themanufacture of pharmaceutical compositions and such compositions cancontain one or more agents selected from the group consisting ofsweetening agents, flavoring agents, coloring agents and preservingagents in order to provide pharmaceutically elegant and palatablepreparations. Tablets may contain the active ingredient in admixturewith nontoxic pharmaceutically acceptable excipients which are suitablefor the manufacture of tablets. These excipients are, for example, inertdiluents, such as calcium carbonate, sodium carbonate, lactose, calciumphosphate or sodium phosphate; granulating and disintegrating agents,for example, corn starch, or alginic acid; binding agents, for example,starch, gelatin or acacia; and lubricating agents, for example magnesiumstearate, stearic acid or talc. The tablets are uncoated or coated byknown techniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate can be employed. Formulations fororal use can be presented as hard gelatin capsules wherein the activeingredient is mixed with an inert solid diluent, for example, calciumcarbonate, calcium phosphate or kaolin, or as soft gelatin capsuleswherein the active ingredient is mixed with water or an oil medium, forexample, peanut oil, liquid paraffin or olive oil.

Certain injectable compositions are aqueous isotonic solutions orsuspensions, and suppositories are advantageously prepared from fattyemulsions or suspensions. Said compositions may be sterilized and/orcontain adjuvants, such as preserving, stabilizing, wetting oremulsifying agents, solution promoters, salts for regulating the osmoticpressure and/or buffers. In addition, they may also contain othertherapeutically valuable substances. Said compositions are preparedaccording to conventional mixing, granulating or coating methods,respectively, and contain about 0.1-75%, or contain about 1-50%, of theactive ingredient.

Suitable compositions for transdermal application include an effectiveamount of a compound of the invention with a suitable carrier. Carrierssuitable for transdermal delivery include absorbable pharmacologicallyacceptable solvents to assist passage through the skin of the host. Forexample, transdermal devices are in the form of a bandage comprising abacking member, a reservoir containing the compound optionally withcarriers, optionally a rate controlling barrier to deliver the compoundof the skin of the host at a controlled and predetermined rate over aprolonged period of time, and means to secure the device to the skin.

Suitable compositions for topical application, e.g., to the skin andeyes, include aqueous solutions, suspensions, ointments, creams, gels orsprayable formulations, e.g., for delivery by aerosol or the like. Suchtopical delivery systems will in particular be appropriate for dermalapplication, e.g., for the treatment of skin cancer, e.g., forprophylactic use in sun creams, lotions, sprays and the like. They arethus particularly suited for use in topical, including cosmetic,formulations well-known in the art. Such may contain solubilizers,stabilizers, tonicity enhancing agents, buffers and preservatives.

As used herein, a topical application may also pertain to an inhalationor to an intranasal application. They may be conveniently delivered inthe form of a dry powder (either alone, as a mixture, for example a dryblend with lactose, or a mixed component particle, for example withphospholipids) from a dry powder inhaler or an aerosol spraypresentation from a pressurized container, pump, spray, atomizer ornebulizer, with or without the use of a suitable propellant.

Dosage forms for the topical or transdermal administration of a compoundof this invention include powders, sprays, ointments, pastes, creams,lotions, gels, solutions, patches and inhalants. The active compound maybe mixed under sterile conditions with a pharmaceutically acceptablecarrier, and with any preservatives, buffers, or propellants that may bedesirable.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients, such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a compound of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane.

Transdermal patches have the added advantage of providing controlleddelivery of a compound of the present invention to the body. Such dosageforms can be made by dissolving or dispersing the compound in the propermedium. Absorption enhancers can also be used to increase the flux ofthe compound across the skin. The rate of such flux can be controlled byeither providing a rate controlling membrane or dispersing the activecompound in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention.

The present invention further provides anhydrous pharmaceuticalcompositions and dosage forms comprising the compounds of the presentinvention as active ingredients, since water may facilitate thedegradation of certain compounds. Anhydrous pharmaceutical compositionsand dosage forms of the invention can be prepared using anhydrous or lowmoisture containing ingredients and low moisture or low humidityconditions. An anhydrous pharmaceutical composition may be prepared andstored such that its anhydrous nature is maintained. Accordingly,anhydrous compositions are packaged using materials known to preventexposure to water such that they can be included in suitable formularykits. Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e. g.,vials), blister packs, and strip packs.

The invention further provides pharmaceutical compositions and dosageforms that comprise one or more agents that reduce the rate by which thecompound of the present invention as an active ingredient willdecompose. Such agents, which are referred to herein as “stabilizers,”include, but are not limited to, antioxidants such as ascorbic acid, pHbuffers, or salt buffers, etc.

The pharmaceutical composition or combination of the present inventioncan be in unit dosage of about 1-1000 mg of active ingredient(s) for asubject of about 50-70 kg, or about 1-500 mg or about 1-250 mg or about1-150 mg or about 0.5-100 mg, or about 1-50 mg of active ingredients.The therapeutically effective dosage of a compound, the pharmaceuticalcomposition, or the combinations thereof, is dependent on the species ofthe subject, the body weight, age and individual condition, the disorderor disease or the severity thereof being treated. A physician, clinicianor veterinarian of ordinary skill can readily determine the effectiveamount of each of the active ingredients necessary to prevent, treat orinhibit the progress of the disorder or disease.

The above-cited dosage properties are demonstrable in vitro and in vivotests using advantageously mammals, e.g., mice, rats, dogs, monkeys orisolated organs, tissues and preparations thereof. The compounds of thepresent invention can be applied in vitro in the form of solutions,e.g., aqueous solutions, and in vivo either enterally, parenterally,advantageously intravenously, e.g., as a suspension or in aqueoussolution. The dosage in vitro may range between about 10-3 molar and10-9 molar concentrations. A therapeutically effective amount in vivomay range depending on the route of administration, between about0.1-500 mg/kg, or between about 1-100 mg/kg.

The compound of the present invention may be administered eithersimultaneously with, or before or after, one or more other therapeuticagent. The compound of the present invention may be administeredseparately, by the same or different route of administration, ortogether in the same pharmaceutical composition as the other agents.

In one embodiment, the invention provides a product comprising acompound of Formula I, II, III, IV or V, and at least one othertherapeutic agent as a combined preparation for simultaneous, separateor sequential use in therapy. In one embodiment, the therapy is thetreatment of a disease or condition mediated by FXR. Products providedas a combined preparation include a composition comprising a compound ofFormula I, II, III, IV or V, and the other therapeutic agent(s) togetherin the same pharmaceutical composition, or the compound of Formula I,II, III, IV or V and the other therapeutic agent(s) in separate form,e.g. in the form of a kit.

In one embodiment, the invention provides a pharmaceutical compositioncomprising a compound of Formula I, II, III, IV or V, and anothertherapeutic agent(s). It is contemplated that the invention provides apharmaceutical composition comprising a compound of Formula I, II, III,IV or V in combination with a naturally occurring non-toxic bile acid,such as ursodeoxycholic acid, as an aid in preventing possible depletionof fat-soluble vitamins secondary to treatment with an FXR agonist.Accordingly, the compounds of the invention may be administeredconcurrently with the naturally occurring non-toxic bile acid, either asseparate entities or as a single formulation comprising a compound ofFormula I, II, III, IV or V, and naturally occurring bile acid.

Optionally, the pharmaceutical composition may comprise apharmaceutically acceptable excipient, as described above.

In one embodiment, the invention provides a kit comprising two or moreseparate pharmaceutical compositions, at least one of which contains acompound of Formula I, II, III, IV or V. In one embodiment, the kitcomprises means for separately retaining said compositions, such as acontainer, divided bottle, or divided foil packet. An example of such akit is a blister pack, as typically used for the packaging of tablets,capsules and the like.

The kit of the invention may be used for administering different dosageforms, for example, oral and parenteral, for administering the separatecompositions at different dosage intervals, or for titrating theseparate compositions against one another. To assist compliance, the kitof the invention typically comprises directions for administration.

In the combination therapies of the invention, the compound of theinvention and the other therapeutic agent may be manufactured and/orformulated by the same or different manufacturers. Moreover, thecompound of the invention and the other therapeutic may be broughttogether into a combination therapy: (i) prior to release of thecombination product to physicians (e.g. in the case of a kit comprisingthe compound of the invention and the other therapeutic agent); (ii) bythe physician themselves (or under the guidance of the physician)shortly before administration; (iii) in the patient themselves, e.g.during sequential administration of the compound of the invention andthe other therapeutic agent.

Accordingly, the invention provides for the use of a compound of FormulaI, II, III, IV or V for treating a disease or condition mediated by FXR,wherein the medicament is prepared for administration, or administeredwith, another therapeutic agent. The invention also provides a compoundof Formula I, II, III, IV or V for use in a method of treating a diseaseor condition mediated by FXR, wherein the compound of Formula I, II,III, IV or V is prepared for administration, or administered with,another therapeutic agent. The invention also provides anothertherapeutic agent for use in a method of treating a disease or conditionmediated by FXR, wherein the other therapeutic agent is prepared foradministration, or administered with, a compound of Formula I, II, III,IV or V.

The invention also provides for the use of a compound of Formula I, II,III, IV or V for treating a disease or condition mediated by FXR,wherein the patient has previously (e.g. within 24 hrs) been treatedwith another therapeutic agent. Alternatively, the invention providesfor the use of another therapeutic agent for treating a disease orcondition mediated by FXR, wherein the patient has previously (e.g.within 24 hrs) been treated with a compound of Formula I, II, III, IV orV.

Additional Embodiments

The present invention further encompasses additional embodimentsdescribed herein.

Embodiment 1

A compound according to formula (I), or a pharmaceutical acceptablesalt, tautomer or stereoisomer thereof,

wherein,

R⁰ is Ring A or C₁₋₆ alkyl; wherein

Ring A is aryl; 5-10 membered heteroaryl comprising 1-3 N, O or Sheteroatoms; 4-6 membered heterocycle comprising 1-2 N, O or Sheteroatoms; or C₃₋₇ cycloalkyl; each of which is unsubstituted orsubstituted by 1-2 substituents each independently represented by R²;wherein L³ and R⁰ can be attached to the same or different ring atoms ofRing A; and the 01-6 alkyl is optionally substituted by 1 to 2 C₁₋₆alkyl;

Ring B is aryl; 5-10 membered heteroaryl comprising 1-3 N, O or Sheteroatoms; 4-6 membered heterocycle comprising 1-2 N, O or Sheteroatoms; or C₃₋₇ cycloalkyl; each of which is unsubstituted orsubstituted by 1-2 substituents each independently represented by R²;

X is (CR⁴R⁵) or C(O);

Y is O, (CR⁴R⁵), *O(CR⁴R⁵) or NR, wherein “*” indicates the point ofattachment of Y to the ring containing the Z ring atoms;

Z¹, Z², Z³, and Z⁴ are each independently CR³ or N;

L¹ is *¹—(CR⁴R⁵)₁₋₂— or *¹—(CR⁴R⁵)—C(O)—NR—, wherein “*¹” indicates thepoint of attachment of L¹ to N;

L² is *²—(CR⁴R⁵)₁₋₂—, *²—(CR⁴R⁵)—C(O), *²—(CR⁴R⁵)—C(O)—NR,*²—(CR⁴R⁵)₂—O—, *²—(CR⁴R⁵)₂—NR—, *²—(CR⁴R⁵)₂—SO₂—, *²—(CR⁴R⁵)₂—NR—C(O)—,or *²—(CR⁴R⁵)—C(O)—NR—(CR⁴R⁵); wherein “*²” indicates the point ofattachment of L² to N;

L³ is —(CR⁴R⁵)— or —C(O)—;

each R² is independently halo, hydroxyl, C₁₋₆ alkyl, or halo-substitutedC₁₋₆ alkyl;

each R³ is independently hydrogen, halo, or C₁₋₆ alkyl; and

R, R⁴ and R⁵ are independently hydrogen or C₁₋₆ alkyl.

Embodiment 2

A compound according to embodiment 1, or a salt, tautomer orstereoisomer thereof, wherein the compound is represented by Formula(II),

wherein

R⁰ is Ring A or C₁₋₆ alkyl; wherein

Ring A is selected from phenyl, pyridyl, pyridazinyl, pyrimidinyl,pyrazinyl and C₃₋₇ cycloalkyl, each of which is unsubstituted orsubstituted by 1-3 substituents each independently represented by R²;wherein L³ and R⁰ can be attached to the same or different ring atoms ofRing A; and the C₁₋₆ alkyl is optionally substituted by 1-2 C₁₋₆ alkyl;

Ring B is selected from phenyl, pyridyl, pyridazinyl, pyrimidinyl,pyrazinyl, 1H-indolyl, and C₃₋₇ cycloalkyl, each of which isunsubstituted or substituted by 1-2 substituents each independentlyrepresented by R²;

X is (CR⁴R⁵);

Y is O, (CR⁴R⁵) or *O(CR⁴R⁵), where “*” indicates the point ofattachment of Y to the ring containing the Z ring atoms;

Z¹, Z², Z³, and Z⁴ are each independently CR³ or N;

L¹ is —(CR⁴R⁵)₁₋₂—,

L² is *²—(CR⁴R⁵)₁₋₂—, *²—(CR⁴R⁵)—C(O)—NR—, *²—(CR⁴R⁵)₂—O—, or*²—(CR⁴R⁵)₂—NR—; where “*²” indicates the point of attachment of L² toN; and

each R² is independently halo, hydroxyl, C₁₋₆ alkyl, or halo-substitutedC₁₋₆ alkyl;

each R³ is independently hydrogen, halo, or C₁₋₆ alkyl;

R, R⁴ and R⁵ are independently hydrogen or C₁₋₆ alkyl.

Embodiment 3

A compound according to embodiment 1 or 2, or a pharmaceuticallyacceptable salt, tautomer or stereoisomer thereof, wherein R⁰ isselected from *³—CH₂C(CH₃)₂—, *³—CH₂CH(CH₃)—, and*³-cyclopropane-1,1-diyl-, wherein “*3” indicates the point ofattachment of R⁰ to L².

Embodiment 4

A compound according to embodiment 1 or 2, or a pharmaceuticallyacceptable salt, tautomer or stereoisomer thereof, wherein the compoundis represented by Formula (III):

wherein

Ring A is phenyl or pyridyl, each of which is unsubstituted orsubstituted by 1-2 substituents each independently represented by R²;

Ring B is selected from phenyl, pyridyl, 1H-indolyl, and cyclopentyl,each of which is unsubstituted or substituted by 1-2 substituents eachindependently represented by R²;

L¹ is —(CR⁴R⁵)—;

L² is selected from *²—(CH₂)—, *²—CH₂C(O)NH—, *²—CH(CH₃)C(O)NH—,*²—CH₂C(O)NHCH₂—, *²—(CH₂)₂O—, and *²—(CH₂)₂NH—; wherein “*2” indicatesthe point of attachment of L² to N;

X is CH₂;

Y is selected from O, CH₂, C(CH₃)₂, and *O—CH₂, wherein “*” indicatesthe point of attachment of Y to the ring containing the Z ring atoms;

Z¹ is CR³ or N;

Z² is CR³;

Z³ is CR³;

Z⁴ is CR³ or N; and

each R² is independently selected from halo, methyl, andtrifluoromethyl;

each R³ is independently hydrogen, halo, or C₁₋₆ alkyl; and

each of —R⁴ and R⁵ is independently hydrogen or methyl.

Embodiment 5

A compound according to any one of embodiments 1 to 4, or apharmaceutically acceptable salt, tautomer or stereoisomer thereof,wherein Y is O.

Embodiment 6

A compound according to any one of embodiments 1 to 5, or apharmaceutically acceptable salt, tautomer or stereoisomer thereof,wherein L¹ is —CH₂—.

Embodiment 7

A compound according to any one of embodiments 1 to 6, or apharmaceutically acceptable salt, tautomer or stereoisomer thereof,wherein L² is selected from *²—(CH₂)—, *²—CH₂C(O)NH—, *²—(CH₂)₂O—, and*²—(CH₂)₂NH—; wherein “*²” indicates the point of attachment of L² to N.

Embodiment 8

A compound according to any one of embodiments 1 to 6, or apharmaceutically acceptable salt, tautomer or stereoisomer thereof,wherein L² is *²—(CH₂)—.

Embodiment 9

A compound according to any one of embodiments 1 to 8, or apharmaceutically acceptable salt, tautomer or stereoisomer thereof,wherein each R² is independently fluoro or methyl.

Embodiment 10

A compound according to any one of embodiments 1 to 9, or apharmaceutically acceptable salt, tautomer or stereoisomer thereof,wherein each R³ is independently selected from hydrogen, fluoro, chloro,and methyl.

Embodiment 11

A compound according to any one of embodiments 1 to 9, or apharmaceutically acceptable salt, tautomer or stereoisomer thereof,wherein

Z¹ is selected from CH, CF, CCH₃, and N;

Z² is selected from CH, CF, CCl, and CCH₃;

Z³ is selected from CH, CF, CCl, and CCH₃; and

Z⁴ is CH or N.

Embodiment 12

A pharmaceutical composition comprising a therapeutically effectiveamount of a compound according to any one of embodiments 1-11 and apharmaceutically acceptable carrier.

Embodiment 13

A combination comprising a therapeutically effective amount of acompound according to any one of embodiments 1-11, and a secondtherapeutic agent being useful in the treatment of cholestasis,intrahepatic cholestasis, estrogen-induced cholestasis, drug-inducedcholestasis, cholestasis of pregnancy, parenteral nutrition-associatedcholestasis, primary biliary cirrhosis (PBC), primary sclerosingcholangitis (PSC), progressive familiar cholestasis (PFIC),non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis(NASH), drug-induced bile duct injury, gallstones, liver cirrhosis,alcohol-induced cirrhosis, cystic fibrosis, bile duct obstruction,cholelithiasis, liver fibrosis, dyslipidemia, atherosclerosis, diabetes,diabetic nephropathy, colitis, newborn jaundice, prevention ofkernicterus, veno-occlusive disease, portal hypertension, metabolicsyndrome, hypercholesterolemia, intestinal bacterial overgrowth, orerectile dysfunction.

Embodiment 14

A method for treating a condition mediated by farnesoid X receptors(FXR) in a subject suffering therefrom, comprising administering to thesubject a therapeutically effective amount of a compound of any one ofembodiments 1-11, or a pharmaceutical composition thereof, andoptionally in combination with a second therapeutic agent.

Embodiment 15

A pharmaceutical composition which comprises a compound according to anyone of embodiments 1-11 for use in the treatment of a condition mediatedby FXR.

Embodiment 16

A compound of Formula (VI)

wherein:

-   -   Ring A is phenyl or pyridyl, each of which is unsubstituted or        substituted by 1-2 substituents each independently represented        by R²;    -   Ring B is selected from phenyl, pyridyl, 1H-indolyl, and        cyclopentyl, each of which is unsubstituted or substituted by        1-2 substituents each independently represented by R²;    -   L¹ is —(CR⁴R⁵)—;    -   L² is selected from *²—(CH₂)—, *²—CH₂C(O)NH—, *²—CH(CH₃)C(O)NH—,        *²—(CH₂)₂O—, and *²—(CH₂)₂NH—; wherein “*2” indicates the point        of attachment of L² to N;    -   X is CH₂;    -   Y is selected from O, CH₂, C(CH₃)₂, and *O—CH₂, wherein “*”        indicates the point of attachment of Y to the ring containing        the Z ring atoms;    -   Z¹ is CR³ or N;    -   Z² is CR³;    -   Z³ is CR³;    -   Z⁴ is CR³ or N;    -   each R² is independently selected from halo, methyl, and        trifluoromethyl;    -   each R³ is independently hydrogen, halo, or C₁₋₆ alkyl;    -   each of R⁴ and R⁵ is independently hydrogen or methyl; and    -   R⁶ is C₁₋₆ alkyl.

Embodiment 17

The compound according to embodiment 16, wherein Y is O.

Embodiment 18

The compound according to embodiment 16, wherein L¹ is —CH₂—.

Embodiment 19

The compound according to embodiment 16, wherein L² is selected from*²—(CH₂)—, *²—CH₂C(O)NH—, *²—(CH₂)₂O—, and *²—(CH₂)₂NH—; wherein “*2”indicates the point of attachment of L² to N; and more particularly,wherein L² is —CH₂—.

Embodiment 20

The compound according to embodiment 16, wherein each R² isindependently fluoro or methyl.

Embodiment 21

The compound according to embodiment 16, wherein each R³ isindependently selected from hydrogen, fluoro, chloro, and methyl.

Embodiment 22

A method for preparing a crystalline form of4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid, comprising the step of reacting4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid with 2-amino-2-hydroxymethyl-propane-1,3-diol ina solvent such as a methanol:dichloromethane solvent (1:1 by volume).

Embodiment 23

A crystalline form of4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid produced according to embodiment 22; and moreparticularly, wherein said crystalline form has a melting point of about125° C. as determined by differential scanning calorimetry.

Embodiment 24

A method for preparing a crystalline form of4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid, comprising the step of: (i) reacting4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid in a solvent such as methanol with aqueousL-arginine; and (ii) optionally, further crystallizing the solidobtained from (i) in a solvent such as an acetonitrile: methanol solvent(2:1 by volume).

Embodiment 25

A crystalline form of4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid produced according to embodiment 24; and moreparticularly, wherein said crystalline form has a melting point of about206° C. as determined by differential scanning calorimetry.

Embodiment 26

A method for preparing a crystalline form of4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid, comprising the step of: (i) reacting4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid in a solvent such as methanol with aqueousL-lysine; and (ii) optionally, further crystallizing the solid obtainedin (i) in a solvent such as acetonitrile.

Embodiment 27

A crystalline form of4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid produced according to embodiment 26.

Embodiment 28

A method for preparing a crystalline form of4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid, comprising the step of reacting4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid with 2-amino-2-hydroxymethyl-propane-1,3-diol in a solvent such asmethanol.

Embodiment 29

A crystalline form of4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid produced according to embodiment 28; and more particularly, whereinsaid crystalline form has a melting point of about 160° C. as determinedby differential scanning calorimetry.

Embodiment 30

A method for preparing a crystalline form of4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid, comprising the step of: (i) reacting4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid in a solvent such as methanol with aqueous L-arginine; and (ii)optionally, further crystallizing the solid obtained from (i) in asolvent such as acetonitrile.

Embodiment 31

A crystalline form of4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid produced according to embodiment 30; and more particularly, whereinsaid crystalline form has a melting point of about 161° C. as determinedby differential scanning calorimetry.

Embodiment 32

A method for preparing a crystalline form of4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid, comprising the step of reacting4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid in a solvent such as methanol with aqueous L-arginine.

Embodiment 33

A crystalline form of4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid produced according to embodiment 32; and more particularly, whereinsaid crystalline form has a melting point of about 206° C. as determinedby differential scanning calorimetry.

Embodiment 34

A method for preparing a crystalline form of4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid, comprising the step of: (i) reacting4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid in a solvent such as methanol with aqueous sodium hydroxide; and(ii) optionally, further crystallizing the solid obtained in (i) in asolvent such as acetonitrile.

Embodiment 35

A crystalline form of4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid produced according to embodiment 34; and more particularly, whereinsaid crystalline form has a melting point of about 161° C. as determinedby differential scanning calorimetry.

Embodiment 36

A method for preparing a crystalline form of4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid, comprising the step of reacting4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid with 2-amino-2-hydroxymethyl-propane-1,3-diol in a solvent such asmethanol.

Embodiment 37

A crystalline form of4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid produced according to embodiment 36; and more particularly, whereinsaid crystalline form has a melting point of about 195.6° C. asdetermined by differential scanning calorimetry.

Embodiment 38

A method for preparing a crystalline form of4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid, comprising the step of: (i) reacting4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid in a solvent such as acetone with aqueous meglumine; and (ii)optionally, further comprising heating the solid obtained in (i) at atemperature ranging from 60-90° C. (e.g., about 80° C.).

Embodiment 39

A crystalline form of4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid produced according to embodiment 38, step (i) having a dehydrationpoint at about 71° C. as determined by differential scanningcalorimetry; or obtained upon further heating in step (ii) having amelting point of about 167.5° C. as determined by differential scanningcalorimetry.

Embodiment 40

A method for preparing a crystalline form of4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid, comprising the step of: (i) reacting4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid and meglumine in a solvent such as methanol; and (ii) furthercomprising heating the reactants at a temperature ranging from 60-90° C.

Embodiment 41

A crystalline form of4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid produced according to embodiment 40; and more particularly, whereinthe crystalline form has a melting point of about 180.6° C. asdetermined by differential scanning calorimetry.

Processes for Making Compounds of the Invention

The compounds of the present invention may be prepared by the routesdescribed in the following schemes or in the Examples. All methodsdescribed herein can be performed in any suitable order unless otherwiseindicated herein or otherwise clearly contradicted by context.

Compounds of Formula (I) wherein L¹ is methyl, L² is *²—CH₂C(O)NH—, L³is C(O) and R⁰ is Ring A may be prepared according to Scheme 1:

Compounds of Formula (I) wherein L¹ is methyl, L² is *²—CH₂C(O)NH—, L³is C(O) and R⁰ is Ring A may be prepared according to Scheme 2:

Compounds of Formula (I) wherein L¹ is methyl, L² is *²—(CH₂)₂NH—, L³ isC(O) and R⁰ is Ring A may be prepared according to Scheme 3:

Compounds of Formula (I) wherein L¹ is methyl, L² is *²—(CH₂)₂O—, L³ isC(O) and R⁰ is Ring A may be prepared according to Scheme 4:

Compounds of Formula (I) wherein L¹ is methyl, L² is *²—CH₂C(O)NH—, L³is C(O) and R⁰ is C₁₋₆ alkyl may be prepared according to Scheme 5:

Compounds of Formula (I) wherein L¹ is methyl, L² is*²—(CH₂)—C(O)NH(CH₂), L³ is C(O) and R⁰ is Ring A may be preparedaccording to Scheme 6:

In each of the above Schemes 1-6, X, Y, Z¹, Z², Z³, Z⁴, A and B are asdefined in any of the above embodiments. Generally, the tricyclic core(VI) is coupled to an amide side chain (Vila, VIIb, VIII, VIId or VIIe)with or without the use of a suitable amide coupling agent such as HATU;followed by hydrolysis to provide a compound of Formula (I).

In each of the above Schemes 1-5, the tricyclic core (VI) may beprepared according to Scheme 7 wherein X, Y, Z¹, Z², Z³ and Z⁴ are asdefined in any of the above embodiments. Typical conditions areexemplified in the syntheses of ethyl2-(6-chloro-4-oxochroman-3-yl)-2-oxoacetate (I-1), ethyl8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate (I-3)and 8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylicacid (I-4), infra.

In the above Scheme 1, the amide side chain (Vila) may be preparedaccording to Scheme 8, wherein A and B are as defined in any of theabove embodiments. Typical conditions are exemplified in the synthesisof methyl 4-fluoro-3-(2-((3-fluorobenzyl)amino) acetamido)benzoate(I-16), infra.

In the above Scheme 2, the amide side chain (VIIb) may be preparedaccording to Scheme 9, wherein A and B are as defined in any of theabove embodiments. Typical conditions are exemplified in the synthesisof methyl 4-((benzylamino)methyl)benzoate (I-23), infra.

In the above Scheme 3, the amide side chain (VIII) may be preparedaccording to Scheme 10, wherein A and B are as defined in any of theabove embodiments. Typical conditions are exemplified in the synthesisof methyl 3-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-4-fluorobenzoate (I-44), methyl3-((2-aminoethyl)amino)-4-fluorobenzoate (I-45) and methyl4-fluoro-3-((2-((3-fluorobenzyl)amino) ethyl)amino)benzoate (I-46),infra.

In the above Scheme 4, the amide side chain (VIId) may be preparedaccording to Scheme 11, wherein B is as defined in any of the aboveembodiments. Typical conditions are exemplified in the synthesis of2-((2-fluorobenzyl)amino)ethanol (I-72), infra.

Each reaction step can be carried out in a manner known to those skilledin the art. For example, a reaction can be carried in the presence of asuitable solvent or diluent or of mixture thereof. A reaction can alsobe carried, if needed, in the presence of an acid or a base, withcooling or heating, for example in a temperature range fromapproximately −30° C. to approximately 150° C. In particular examples, areaction is carried in a temperature range from approximately 0° C. to100° C., and more particularly, in a temperature range from roomtemperature to approximately 80° C., in an open or closed reactionvessel and/or in the atmosphere of an inert gas, for example nitrogen.

The invention also relates to those forms of the process in which acompound obtainable as an intermediate at any stage of the process isused as starting material and the remaining process steps are carriedout, or in which a starting material is formed under the reactionconditions or is used in the form of a derivative, for example in aprotected form or in the form of a salt, or a compound obtainable by theprocess according to the invention is produced under the processconditions and processed further in situ. Compounds of the invention andintermediates can also be converted into each other according to methodsgenerally known to those skilled in the art. Intermediates and finalproducts can be worked up and/or purified according to standard methods,e.g. using chromatographic methods, distribution methods, (re-)crystallization, and the like.

In the reactions described, reactive functional groups, for examplehydroxyl, amino, imino, thio or carboxy groups, where these are desiredin the final product, may be protected to avoid their unwantedparticipation in the reactions. A characteristic of protecting groups isthat they can be removed readily (i.e. without the occurrence ofundesired secondary reactions) for example by solvolysis, reduction,photolysis or alternatively under physiological conditions (e.g. byenzymatic cleavage). Conventional protecting groups may be used inaccordance with standard practice (see e.g., T.W. Greene and P. G. M.Wuts in “Protective Groups in Organic Chemistry,” 4^(th) Ed.,Wiley-Interscience, 2006, and subsequent versions thereof).

All the above-mentioned process steps mentioned herein before andhereinafter can be carried out under reaction conditions that are knownto those skilled in the art, including those mentioned specifically, inthe absence or, customarily, in the presence of solvents or diluents,including, for example, solvents or diluents that are inert towards thereagents used and dissolve them, in the absence or presence ofcatalysts, condensation or neutralizing agents, for example ionexchangers, such as cation exchangers, e.g. in the H⁺ form, depending onthe nature of the reaction and/or of the reactants at reduced, normal orelevated temperature, for example in a temperature range of from about−100° C. to about 190° C., including, for example, from approximately−80° C. to approximately 150° C., for example at from −80 to −60° C., atroom temperature, at from −20 to 40° C. or at reflux temperature, underatmospheric pressure or in a closed vessel, where appropriate underpressure, and/or in an inert atmosphere, for example under an argon ornitrogen atmosphere.

At all stages of the reactions, mixtures of isomers that are formed canbe separated into the individual isomers, for example diastereoisomersor enantiomers, or into any desired mixtures of isomers, for exampleracemates or mixtures of diastereoisomers. Mixtures of isomersobtainable according to the invention can be separated in a manner knownto those skilled in the art into the individual isomers;diastereoisomers can be separated, for example, by partitioning betweenpolyphasic solvent mixtures, recrystallization and/or chromatographicseparation, for example over silica gel or by e.g. medium pressureliquid chromatography over a reversed phase column, and racemates can beseparated, for example, by the formation of salts with optically puresalt-forming reagents and separation of the mixture of diastereoisomersso obtainable, for example by means of fractional crystallization, or bychromatography over optically active column materials.

The solvents from which those solvents that are suitable for anyparticular reaction may be selected include those mentioned specificallyor, for example, water, esters, such as lower alkyl-lower alkanoates,for example ethyl acetate, ethers, such as aliphatic ethers, for examplediethyl ether, or cyclic ethers, for example tetrahydrofuran or dioxane,liquid aromatic hydrocarbons, such as benzene or toluene, alcohols, suchas methanol, ethanol or 1- or 2-propanol, nitriles, such asacetonitrile, halogenated hydrocarbons, such as methylene chloride orchloroform, acid amides, such as dimethylformamide or dimethylacetamide, bases, such as heterocyclic or heteroaromatic nitrogen bases,for example pyridine or N-methylpyrrolidin-2-one, carboxylic acidanhydrides, such as lower alkanoic acid anhydrides, for example aceticanhydride, cyclic, linear or branched hydrocarbons, such as cyclohexane,hexane or isopentane, methylcyclohexane, or mixtures of those solvents,for example aqueous solutions, unless otherwise indicated in thedescription of the processes. Such solvent mixtures may also be used inworking up, for example by chromatography or partitioning.

The compounds of the present invention are either obtained in the freeform, as a salt thereof, or as prodrug derivatives thereof. When both abasic group and an acid group are present in the same molecule, thecompounds of the present invention may also form internal salts, e.g.,zwitterionic molecules. In many cases, the compounds of the presentinvention are capable of forming acid and/or base salts by virtue of thepresence of amino and/or carboxyl groups or groups similar thereto. Asused herein, the terms “salt” or “salts” refers to an acid addition orbase addition salt of a compound of the invention. “Salts” include inparticular “pharmaceutical acceptable salts”. The term “pharmaceuticallyacceptable salts” refers to salts that retain the biologicaleffectiveness and properties of the compounds of this invention and,which typically are not biologically or otherwise undesirable.

Salts of compounds of the present invention having at least onesalt-forming group may be prepared in a manner known to those skilled inthe art. For example, salts of compounds of the present invention havingacid groups may be formed, for example, by treating the compounds withmetal compounds, such as alkali metal salts of suitable organiccarboxylic acids, e.g. the sodium salt of 2-ethylhexanoic acid, withorganic alkali metal or alkaline earth metal compounds, such as thecorresponding hydroxides, carbonates or hydrogen carbonates, such assodium or potassium hydroxide, carbonate or hydrogen carbonate, withcorresponding calcium compounds or with ammonia or a suitable organicamine, stoichiometric amounts or only a small excess of the salt-formingagent preferably being used. Acid addition salts of compounds of thepresent invention are obtained in customary manner, e.g. by treating thecompounds with an acid or a suitable anion exchange reagent. Internalsalts of compounds of the present invention containing acid and basicsalt-forming groups, e.g. a free carboxy group and a free amino group,may be formed, e.g. by the neutralization of salts, such as acidaddition salts, to the isoelectric point, e.g. with weak bases, or bytreatment with ion exchangers. Salts can be converted into the freecompounds in accordance with methods known to those skilled in the art.Metal and ammonium salts can be converted, for example, by treatmentwith suitable acids, and acid addition salts, for example, by treatmentwith a suitable basic agent.

Pharmaceutically acceptable acid addition salts can be formed withinorganic acids and organic acids, e.g., acetate, aspartate, benzoate,besylate, bromide/hydrobromide, bicarbonate/carbonate,bisulfate/sulfate, camphorsulfonate, chloride/hydrochloride,chlorotheophyllinate, citrate, ethandisulfonate, fumarate, gluceptate,gluconate, glucuronate, hippurate, hydroiodide/iodide, isethionate,lactate, lactobionate, laurylsulfate, malate, maleate, malonate,mandelate, mesylate, methylsulphate, naphthoate, napsylate, nicotinate,nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate,phosphate/hydrogen phosphate/dihydrogen phosphate, polygalacturonate,propionate, stearate, succinate, sulfosalicylate, tartrate, tosylate andtrifluoroacetate salts.

Inorganic acids from which salts can be derived include, for example,hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like.

Organic acids from which salts can be derived include, for example,acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid,malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,toluenesulfonic acid, sulfosalicylic acid, and the like.Pharmaceutically acceptable base addition salts can be formed withinorganic and organic bases.

Inorganic bases from which salts can be derived include, for example,ammonium salts and metals from columns I to XII of the periodic table.In certain embodiments, the salts are derived from sodium, potassium,ammonium, calcium, magnesium, iron, silver, zinc, and copper;particularly suitable salts include ammonium, potassium, sodium, calciumand magnesium salts.

Organic bases from which salts can be derived include, for example,primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines, basic ionexchange resins, and the like. Certain organic amines includeisopropylamine, benzathine, cholinate, diethanolamine, diethylamine,lysine, meglumine, piperazine and tromethamine.

Other pharmaceutically acceptable salts can be derived from L-arginine,TRIS (2-amino-2-hydroxymethyl-1,3-propanediol), adipic acid (adipate),L-ascorbic acid (ascorbate), capric acid (caprate), sebacic acid(sebacate), 1-hydroxy-2-naphthoic acid (xinafoate), L-glutamic acid(glutamate), glutaric acid (glutarate), triphenylacetic acid(trifenatate) and galactaric acid/mucic acid (mucate).

The pharmaceutically acceptable salts of the present invention can besynthesized from a parent compound, a basic or acidic moiety, byconventional chemical methods. Generally, such salts can be prepared byreacting free acid forms of these compounds with a stoichiometric amountof the appropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate,bicarbonate or the like), or by reacting free base forms of thesecompounds with a stoichiometric amount of the appropriate acid. Suchreactions are typically carried out in water or in an organic solvent,or in a mixture of the two. Generally, use of non-aqueous media likeether, ethyl acetate, ethanol, isopropanol, or acetonitrile isdesirable, where practicable. Lists of additional suitable salts can befound, e.g., in “Remington: The Science and Practice of Pharmacy,”21^(st) Ed., Pharmaceutical Press 2011; and in “Pharmaceutical Salts:Properties, Selection, and Use” by Stahl and Wermuth (2^(nd) Rev. Ed.,Wiley-VCH 2011, and subsequent versions thereof).

The present invention also provides pro-drugs of the compounds of thepresent invention that converts in vivo to the compounds of the presentinvention. A pro-drug is an active or inactive compound that is modifiedchemically through in vivo physiological action, such as hydrolysis,metabolism and the like, into a compound of this invention followingadministration of the prodrug to a subject. The suitability andtechniques involved in making and using pro-drugs are well known bythose skilled in the art. Prodrugs can be conceptually divided into twonon-exclusive categories, bioprecursor prodrugs and carrier prodrugs.(See, The Practice of Medicinal Chemistry, Ch. 31-32 (Ed. Wermuth,Academic Press, San Diego, Calif., 2001, and subsequent versionsthereof). Generally, bioprecursor prodrugs are compounds, which areinactive or have low activity compared to the corresponding active drugcompound that contain one or more protective groups and are converted toan active form by metabolism or solvolysis. Both the active drug formand any released metabolic products should have acceptably low toxicity.

Carrier prodrugs are drug compounds that contain a transport moiety,e.g., that improve uptake and/or localized delivery to a site(s) ofaction. Desirably for such a carrier prodrug, the linkage between thedrug moiety and the transport moiety is a covalent bond, the prodrug isinactive or less active than the drug compound, and any releasedtransport moiety is acceptably non-toxic. For prodrugs where thetransport moiety is intended to enhance uptake, typically the release ofthe transport moiety should be rapid. In other cases, it is desirable toutilize a moiety that provides slow release, e.g., certain polymers orother moieties, such as cyclodextrins. Carrier prodrugs can, forexample, be used to improve one or more of the following properties:increased lipophilicity, increased duration of pharmacological effects,increased site-specificity, decreased toxicity and adverse reactions,and/or improvement in drug formulation (e.g., stability, watersolubility, suppression of an undesirable organoleptic or physiochemicalproperty). For example, lipophilicity can be increased by esterificationof (a) hydroxyl groups with lipophilic carboxylic acids (e.g., acarboxylic acid having at least one lipophilic moiety), or (b)carboxylic acid groups with lipophilic alcohols (e.g., an alcohol havingat least one lipophilic moiety, for example aliphatic alcohols).

Exemplary prodrugs are, e.g., esters of free carboxylic acids and S-acylderivatives of thiols and O-acyl derivatives of alcohols or phenols,wherein acyl has a meaning as defined herein. Suitable prodrugs areoften pharmaceutically acceptable ester derivatives convertible bysolvolysis under physiological conditions to the parent carboxylic acid,e.g., alkyl esters, cycloalkyl esters, alkenyl esters, benzyl esters,mono- or di-substituted alkyl esters, such as the ω-(amino, mono- ordi-alkylamino, carboxy, alkoxycarbonyl) alkyl esters, theα-(alkanoyloxy, alkoxycarbonyl or di-alkylaminocarbonyl) alkyl esters,such as the pivaloyloxymethyl ester and the like conventionally used inthe art. In addition, amines have been masked as arylcarbonyloxymethylsubstituted derivatives which are cleaved by esterases in vivo releasingthe free drug and formaldehyde (Bundgaard, J. Med. Chem. 2503 (1989),and subsequent versions thereof). Moreover, drugs containing an acidicNH group, such as imidazole, imide, indole and the like, have beenmasked with N-acyloxymethyl groups (Bundgaard, Design of Prodrugs,Elsevier (1985), and subsequent versions thereof). Hydroxy groups havebeen masked as esters and ethers. EP 039,051 (Sloan and Little)discloses Mannich-base hydroxamic acid prodrugs, their preparation anduse.

Furthermore, the compounds of the present invention, including theirsalts, may also be obtained in the form of hydrates, or their crystalsmay, for example, include the solvent used for crystallization.Different crystalline forms may be present. The compounds of the presentinvention may inherently or by design form solvates withpharmaceutically acceptable solvents (including water); therefore, it isintended that the invention embrace both solvated and unsolvated forms.The term “solvate” refers to a molecular complex of a compound of thepresent invention (including pharmaceutically acceptable salts thereof)with one or more solvent molecules. Such solvent molecules are thosecommonly used in the pharmaceutical art, which are known to be innocuousto the recipient, e.g., water, ethanol, and the like. The term “hydrate”refers to the complex where the solvent molecule is water. The compoundsof the present invention, including salts, hydrates and solvatesthereof, may inherently or by design form polymorphs.

Compounds of the invention in unoxidized form may be prepared fromN-oxides of compounds of the invention by treating with a reducing agent(e.g., sulfur, sulfur dioxide, triphenyl phosphine, lithium borohydride,sodium borohydride, phosphorus trichloride, tribromide, or the like) ina suitable inert organic solvent (e.g. acetonitrile, ethanol, aqueousdioxane, or the like) at 0 to 80° C.

EXAMPLES

The examples provided herein are offered to illustrate but not limit thecompounds of the invention, as well as the preparation of such compoundsand intermediates. It is understood that if there appears to be adiscrepancy between the name and structure of a particular compound, thestructure is to be considered correct as the compound names weregenerated from the structures. All the variables are as defined herein.

All starting materials, building blocks, reagents, acids, bases,dehydrating agents, solvents and catalysts utilized to synthesize thecompounds of the present invention are either commercially available orcan be produced by organic synthesis methods known to one of ordinaryskill in the art (Houben-Weyl Science of Synthesis volumes 1-48, GeorgThieme Verlag, and subsequent versions thereof). Furthermore, thecompounds of the present invention can be produced by organic synthesismethods known to one of ordinary skill in the art as shown in thefollowing examples.

Temperatures are given in degrees Celsius. If not mentioned otherwise,all evaporations are performed under reduced pressure, typically betweenabout 15 mm Hg and 100 mm Hg (20-133 mbar). The structure of finalproducts, intermediates and starting materials is confirmed by standardanalytical methods, e.g., microanalysis and spectroscopiccharacteristics, e.g., MS, IR, NMR. Abbreviations used are thoseconventional in the art.

Unless mentioned otherwise, melting points were calculated bydifferential scanning calorimetry (DSC) using TA Q2000 or TA Discoverydifferential scanning calorimeters at a scanning rate of 10° C./min. Theaccuracy of the measured sample temperature is generally within about±1° C.

NMR and LC-MS methodologies are well-known in the art. The methodsdescribed herein are merely illustrative, and are not consideredlimiting.

NMR. NMR spectra were recorded on either a Bruker AVANCE-400 operatingat a proton frequency of 400.13 MHz equipped with a 5 mm QNP cryoprobe(¹H/¹³C/¹⁹F/³¹P); or a Bruker AVANCE-600 spectrometer operating at afrequency of 600.13 MHz equipped with a 5 mm Z-gradient TCI cryoprobe ora 5-mm TXI cryoprobe. Unless otherwise indicated, samples were acquiredat a temperature of 300° K, and spectra were referenced to theappropriate solvent peak.LC-MS Methods. Mass spectra were acquired on LC-MS systems usingelectrospray, chemical and electron impact ionization methods from arange of instruments. Typical methods are described below.

Method 1:

Waters Acquity Binary Gradient Pump; Waters Acquity PDA Detector. WatersAuto sampler; Waters Quattro micro API Mass Spectrometer with ESI andAPCI ion source; UPLC Column: Waters Acquity; BEH; C18 1.7 um 50×2.1 mm;Mobile Phase: (A) H₂O+0.025% TFA and (B) Acetonitrile+0.025% TFA.Gradient: 0.4 mL/min, initial 15% B ramp to 95% B over 3.0 mins, thenhold until 4.0 mins, return to 15% B at 4.1 mins until end of run, thenequilibrated the column for 2.0 mins. MS Scan: 100 to 1000 amu in 0.5seconds per channel; Diode Array Detector: 200 nm and 400 nm.

Method 2:

Waters Acquity Binary Gradient Pump; Waters Acquity PDA Detector. WatersAuto sampler; Waters Quattro micro API Mass Spectrometer with ESI andAPCI ion source; UPLC Column: Waters Acquity; BEH; C18 1.7 um 50×2.1 mm;Mobile Phase: (A) H₂O+0.025% TFA and (B) Acetonitrile+0.025% TFA.Gradient: 0.4 mL/min, initial 20% B ramp to 90% B over 2.0 mins, thenhold until 4.0 mins, return to 20% B at 4.1 mins until end of run, thenequilibrated the column for 2.0 mins. MS Scan: 100 to 1000 amu in 0.5seconds per channel; Diode Array Detector: 200 nm and 400 nm.

Method 3:

Agilent 1200 sl/6140 system; UPLC Column: Waters Acquity; HSS T3; C181.8 um 50×2.0 mm; Mobile Phase: (A) H₂O+0.05% TFA and (B)Acetonitrile+0.035% TFA. Gradient: 0.9 mL/min, initial 10% B ramp to100% B over 1.95 mins, then return to 10% B at 2.00 mins until end ofrun. MS Scan: 100 to 1000 amu in 0.5 seconds per channel; Diode ArrayDetector: 190 nm and 400 nm; Drift tube temperature: 50° C. and N₂ gasflow: 40 Psi for ELSD Detector.

Method 4:

Agilent 1100 sl/1946 system; UPLC Column: Waters Atlantis; C18 1.8 um50×2.0 mm; Mobile Phase: (A) H₂O+0.05% TFA and (B) Acetonitrile+0.035%TFA. Gradient: 1.0 mL/min, initial 10% B ramp to 90% B over 3.00 mins,then return to 10% B at 3.5 mins until end of run. MS Scan: 100 to 1000amu in 0.5 seconds per channel; Diode Array Detector: 190 nm and 400 nm;Drift tube temperature: 50° C. and N₂ gas flow: 40 Psi for ELSDDetector.

Analytical method: WATERS ZQ SHIMADZU LEAP CTC, ZORBAX SB-C8 30*4.6 mm,3.5 um, UV1:220 nm, UV 2:254 nm, A:H₂O (0.03% TFA), B:CH₃CN (0.05% TFA),Flow: 2.000 (ml/min), Time/% B: 0/5, 1.90/95, 2.30/95, 2.31/5, 2.50/5.

Abbreviations

Boc tertiary butyl carboxybr broadd doubletdd doublet of doubletsDIAD diisopropyl azodicarboxylateDIEA diethylisopropylamine

DMF N,N-dimethylformamide

DMSO dimethylsulfoxideESI electrospray ionizationEtOAc ethyl acetateHATU 2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphatehr(s) hour(s)HPLC high pressure liquid chromatographyLCMS liquid chromatography and mass spectrometryMeOH methanolMS mass spectrometrym multipletmg milligrammin(s) minute(s)ml millilitermmol millimolm/z mass to charge ratioNaHDMS sodium bis(trimethylsilyl)amide

NMM N-methylmorpholine

NMR nuclear magnetic resonancePPh₃ triphenylphosphiner.t. retention times singlettripletTFA trifluoroacetic acidTHF tetrahydrofuranTRIS (2-amino-2-hydroxymethyl-1,3-propanediol)Tris.HCl aminotris(hydroxymethyl)methane hydrochloride

Intermediates Ethyl 2-(6-chloro-4-oxochroman-3-yl)-2-oxoacetate (I-1)

A solution of 6-chloro-2,3-dihydrochromen-4-one (109 mmol) in THF wastreated with a solution of NaHMDS (60 mL, 120 mmol, 1.1 eq., 2M in THF)in THF at −78° C. under nitrogen. After stirring for 30 min, diethyloxalate (22 mL, 163 mmol, 1.5 eq.) was added at −78° C. dropwise andthen stirred for 1 hr at room temperature. Subsequently the reaction wasquenched with 1N HCl until the pH value was adjusted to 3. The resultingmixture was extracted with EtOAc (200 mL×3). The combined organic phasewas washed with brine and dried over Na₂SO₄. The solvent was removed invacuo to give I-1 as a yellow solid. MS (m/z): 283 (M+H)⁺.

Ethyl 8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate(I-3)

To a warm solution of I-1 (109 mmol) in ethanol was added1-methylhydrazine (109 mmol,) and the solution was stirred for 12 hrs atroom temperature. After the solvent was removed in vacuo, the residuewas purified by column chromatography (petroleum ether: ethylacetate=95:5) to give I-2 and the desired product I-3 as a yellow solid.¹H-NMR: (300 MHz, CDCl₃): δ 7.69 (d, J=2.7 Hz, 1H), 7.13 (dd, J=2.4 Hz,J=8.7 Hz, 1H), 6.86 (d, J=8.7 Hz, 1H), 5.43 (s, 2H), 4.38 (q, J=7.2 Hz,2H), 4.22 (s, 3H), 1.41 (t, J=7.2 Hz, 3H). MS (m/z): 293 (M+H)⁺.

8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-4)

To a solution of I-3 (44 mmol) in 100 mL of THF/water (4:1) was addedNaOH (89 mmol, 2 eq.) and the mixture was heated at 60° C. for 8 hrs.Subsequently the resulting white solid was filtrated and washed withmethanol and the solid was treated with 100 mL of 1 N HCl. The resultingsolid was collected by filtration, washed with methanol thoroughly anddried in vacuum to give I-4 as a white solid. ¹H-NMR (300 MHz, CDCl₃): δ13.05 (brs, 1H), 7.72 (d, J=2.7 Hz, 1H), 7.34 (dd, J=2.7 Hz, J=8.7 Hz,1H), 7.06 (d, J=8.7 Hz, 1H), 5.41 (s, 2H), 4.19 (s, 3H). MS (m/z): 265(M+H)⁺.

The following intermediates were prepared according to the proceduresdescribed for the synthesis of I-4 using the appropriate reagents.

Compound Structure Characterization Data I-5 

¹H NMR (400 MHz, DMSO): δ 7.57 (dd, J = 3.0, 9.3 Hz, 1H), 7.15 (td, J =3.0, 8.7 Hz, 1H), 7.06 (dd, J = 4.9, 9.0 Hz, 1H), 5.36 (s, 2H), 4.18 (s,3H). I-6 

¹H NMR (400 MHz, DMSO): δ 7.82 (dd, J = 8.8, 11.3 Hz, 1H), 7.23 (dd, J =7.2, 11.8 Hz, 1H), 5.38 (s, 2H), 4.16 (s, 3H). I-7 

Commercial I-8 

MS (m/z): 245 (M + H)⁺ I-9 

¹H NMR (400 MHz, DMSO): δ 7.79-7.69 (m, 1H), 7.19-7.09 (m, 2H), 5.43 (s,2H), 4.16 (s, 3H). I-10

¹H NMR (400 MHz, DMSO): δ 7.81-7.73 (m, 1H), 6.98-6.90 (m, 2H), 5.42 (s,2H), 4.16 (s, 3H). I-11

MS (m/z): 245 (M + H)⁺ I-12

¹H NMR (400 MHz, DMSO) δ 7.47 (ddd, J = 9.1, 2.9, 1.7 Hz, 1H), 7.38(ddd, J = 11.0, 9.0, 2.9 Hz, 1H), 5.46 (s, 2H), 4.19 (s, 3H). I-13

¹H NMR (400 MHz, DMSO): δ 7.69 (s, 1H), 7.06 (d, J = 0.4 Hz, 1H), 5.38(s, 2H), 4.17 (s, 3H), 2.30 (s, 3H). I-14

¹H NMR (400 MHz, DMSO): δ 7.51 (d, J = 2.4, 1H), 7.23 (d, J = 2.1 Hz,1H), 5.40 (s, 2H), 4.10 (s, 3H), 2.18 (s, 3H). I-15

¹H NMR (400 MHz, DMSO): δ 7.53-7.50 (m, 1H), 7.44 (dd, J = 2.3, 10.6 Hz,1H), 5.47 (s, 2H), 4.10 (s, 3H).

Methyl 4-fluoro-3-(2-((3-fluorobenzyl)amino)acetamido)benzoate (I-16)

Bromoacetyl bromide (1.1 mL, 12.5 mmol) was added dropwise to a mixtureof the methyl 3-amino-4-fluorobenzoate (10.9 mmol) anddiisopropylethylamine (2.8 mL, 16.4 mmol) in dichloromethane at 0° C.After stirring for 30 mins at 0° C., the reaction mixture was dilutedwith dichloromethane and water. The layers were separated and theaqueous phase was washed with dichloromethane (2×). the combined organiclayers were dried over MgSO₄ and concentrated (aspirator) to give a darkbrown liquid. LCMS showed that the amide was the major component and theaniline was a minor component. The mixture was dissolved in DMF (30 mL)and potassium carbonate (12.3 mmol) was added. To the mixture was added3-fluorobenzylamine (10.9 mmol) and the reaction stirred at roomtemperature for 16 hrs. The reaction was diluted with water (300 mL) andthe aqueous solution was extracted with ethyl acetate (3×). The combinedorganic layers were washed with water, brine, and were dried over MgSO₄.The mixture was filtered and concentrated and the crude residue waspurified by flash chromatography (silica gel, 0-60% ethylacetate/hexanes) to give I-16. ¹H NMR (400 MHz, d₄-MeOH): δ 8.80 (dd,J=8.0, 4.0 Hz, 1H), 7.89-7.85 (m, 1H), 7.56-7.51 (m, 1H), 7.38-7.29 (m,3H), 7.25 (app dt, J=8.3, 6.0 Hz, 1H), 4.35 (s, 2H), 4.09 (s, 2H), 3.91(s, 3H).

The following intermediates were prepared according to the proceduresdescribed for the synthesis of I-16 using the appropriate reagents.

Compound Structure Characterization Data I-17

¹H NMR (400 MHz, CDCl₃): δ 9.70 (br s, 1H), 9.02 (dd, J = 2.2, 7.6 Hz,1H), 7.80 (ddd, J = 2.2, 5.1, 8.6 Hz, 1H), 7.34-7.26 (m, 2H), 7.19- 7.03(m, 3H), 3.93-3.88 (m, 5H), 3.47 (s, 2H). I-18

¹H NMR (400 MHz, CDCl₃): δ 9.41 (s, 1H), 8.05 (t, J = 1.9 Hz, 1H), 8.00(ddd, J = 1.0, 2.2, 8.1 Hz 1H), 7.82-7.78 (m, 1H), 7.46-7.30 (m, 7H),3.95 (s, 3H), 3.89 (s, 2H), 3.47 (s, 2H). MS (m/z): 299.1 (M + H)⁺ I-19

MS (m/z): 317.1 (M + H)⁺ I-20

MS (m/z): 331.1 (M + H)⁺

2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)aceticacid (I-22)

Step 1

To a suspension of the8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-4) (1.9 mmol) in CH₂Cl₂ (10 mL) was added catalytic amount of DMF (25uL) and dropwise oxalyl chloride (0.78 mL, 8.9 mmol) at 0° C. Theresulting suspension was warmed up to room temperature and stirred for 1hr. The solvent was removed under vacuum to dryness completely (need toremove oxalyl chloride completely). The obtained residue was dissolvedwith CH₂Cl₂ (10 mL) and then dropwise into a solution of methyl2-((3-fluorobenzyl)amino)acetate (1.9 mmol) in CH₂Cl₂ (10 mL) in thepresence of DIEA (0.56 mL, 3.8 mmol). The reaction mixture was stirredfor 30 min at room temperature and then was charged 50 mL water. Organiclayer was separated and aqueous layer was extracted with CH₂Cl₂ (50 mL).Combined organic layers were washed with H₂O and brine successively,dried over Na₂SO₄, filtered and concentrated to give the crude productwhich was purified by column chromatography (0-60% EtOAc in hexanes) toyield methyl2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetate(I-21). ¹H NMR (400 MHz, CDCl₃): δ 7.47-7.43 (m, 2H), 7.34-7.29 (m, 1H),7.23-7.19 (m, 1H), 7.18-7.04 (m, 2H), 6.97 (dd, J=2.3, 8.7 Hz, 1H),5.57-4.12 (m, 9H), 3.77-3.74 (m, 3H). Mixture of rotamers. MS (m/z):444.1 (M+H)⁺.

Step 2

Methyl2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetate(I-21) (1.62 mmol) was dissolved in THF/MeOH/H₂O (3:2:1, 10 mL) andfollowed by addition of LiOH monohydrate (0.408 g, 9.72 mmol). Thereaction mixture was stirred at room temperature for 1 hr and dilutedwith 10 ml of water and acidified to pH=2.0. Solid was collected anddried to yield2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)aceticacid (I-22). ¹H NMR (400 MHz, DMSO): δ 12.73 (s, 1H), 7.71 (t, J=2.6 Hz,1H), 7.42-7.30 (m, 3H), 7.24-7.15 (m, 2H), 7.07-7.02 (m, 1H), 5.41-4.10(m, 9H). Mixture of rotamers. MS (m/z) 430.0 (M+H)⁺.

Methyl 4-((benzylamino)methyl)benzoate (I-23)

Benzyl amine (8.65 mL, 79 mmol) (Aldrich) and methyl 4-formylbenzoate(79 mmol) (Aldrich) were dissolved in methanol (Volume: 350 mL) and themixture was stirred for 2 hr at room temperature, resulting in a whiteprecipitate. The reaction mixture was cooled to 0° C. and sodiumborohydride (158 mmol) (Aldrich) was added portion-wise over the courseof 15 min. After stirring for 1 hr at 0° C. the mixture was allowed towarm to room temperature. After stirring for an additional 1 hr themixture was cooled in an ice bath and the reaction was quenched slowlywith water (15 mL, 833 mmol). The resulting mixture was allowed to stirfor 15 min. After removal of solvent (aspirator) the mixture wasdissolved in EtOAc and washed with H₂O. The aqueous phase was washedwith EtOAc and the combined organic layers were dried (MgSO₄). The crudematerial was purified by chromatography (silica gel, loaded neat withDCM rinse, 0-40% DCM/(10% MeOH in DCM)) to give I-23. ¹H NMR (400 MHz,DMSO): δ 9.17 (bs, 2H), 7.95-7.96 (m, 2H), 7.56-7.54 (m, 2H), 7.44-7.34(m, 5H), 4.19 (s, 1H), 4.12 (s, 1H), 3.78 (s, 3H).

The following intermediates were prepared according to the proceduresdescribed for the synthesis of I-23 using the appropriate reagents.

Compound Structure Characterization Data I-24

MS (m/z): 274.2 (M + H)⁺; r.t. = 1.281 I-25

¹H NMR (400 MHz, DMSO): δ 9.42 (br s, 1H, NH), 8.04 (d, J = 8.2 Hz, 2H),7.78 (d, J = 8.2 Hz, 2H), 7.62 (dd, J = 8.0 Hz, 4.0, 1H), 7.44-7.32 (m,3H), 4.36 (br s, 2H), 4.30 (s, 2H), 3.88 (s, 3H). MS (m/z): 274.1 (M +H)⁺; r.t. = 1.38. I-26

MS (m/z): 292.1 (M + H)⁺ I-27

MS (m/z): 270.1 (M + H)⁺ I-28

MS (m/z): 292.1 (M + H)⁺ I-29

MS (m/z): 288.1 (M + H)⁺ I-30

MS (m/z): 288.1 (M + H)⁺ I-31

MS (m/z): 295.2 (M + H)⁺; r.t. = 1.133 I-32

MS (m/z): 275.2 (M + H)⁺; r.t. = 1.147 I-33

MS (m/z): 275.1 (M + H)⁺; r.t. = 1.151 I-34

MS (m/z): 291.1 (M + H)⁺; r.t. = 1.304 I-35

MS (m/z): 292.2 (M + H)⁺; r.t. = 1.088 I-36

MS (m/z): 274.2 (M + H)⁺; r.t. = 1.081 I-37

¹H NMR (400 MHz, DMSO): δ 7.83 (s, 1H), 7.73 (d, J = 8.2 Hz, 2H), 7.32(d, J = 8.2 Hz, 2H), 7.24 (m, 5H), 7.14 (m, 1H), 3.62 (d, J = 19.2 Hz,4H), 3.25 (s, 1H). I-38

¹H NMR (400 MHz, DMSO): δ 9.76 (br s, 1H, NH), 8.06 (d, J = 8.2 Hz, 2H),7.76 (d, J = 8.2 Hz, 2H), 7.62 (t, J = 8.0 Hz, 1H), 7.50 (app q, J = 7.8Hz, 1H), 7.34-7.28 (m, 2H), 4.34 (br s, 2H), 4.28 (s, 2H), 3.80 (s, 3H).MS (m/z): 274.1 (M + H)⁺; r.t. = 0.239. I-39

¹H NMR (400 MHz, d₄-MeOH): δ 7.99 (d, J = 3.2 Hz, 1H), 7.89 (d, J = 8.0Hz, 1H), 7.72 (t, J = 8.0 Hz, 1H), 7.63-7.56 (m, 2H), 7.28-7.20 (m, 2H),4.45 (br s, 2H), 4.38 (s, 2H), 3.91 (s, 3H). MS (m/z): 292.1 (M + H)⁺;r.t. = 1.42. I-40

¹H NMR (400 MHz, DMSO): δ 9.52 (s, 1H), 7.89 (d, J = 8.2 Hz, 2H),7.80-7.64 (m, 2H), 7.28-7.19 (m, 2H), 7.20 (t, J = 7.7 Hz, 1H), 4.40 (brs, 4H), 3.91 (s, 3H). MS (m/z): 292.1 (M + H)⁺; r.t. = 1.52. I-41

¹H NMR (400 MHz, DMSO): δ 7.80 (d, J = 7.2 Hz, 1H), 7.80-7.55 (m, 5H),7.33-7.26 (m, 2H), 4.20 (br s, 2H), 4.14 (s, 2H), 3.90 (s, 3H). MS(m/z): 292.1 (M + H)⁺; r.t. = 1.22. I-42

¹H NMR (400 MHz, d₄-MeOH): δ 9.22 (d, J = 2.2 Hz, 1H), 8.34 (d, J = 8.2Hz, 1H), 7.48-7.40 (m, 2H), 7.33-7.27 (m, 3H). 7.10 (t, J = 8.1 Hz, 1H),4.40 (br s, 2H), 4.34 (s, 2H), 3.94 (s, 3H). MS (m/z): 275.1 (M + H)⁺;r.t. = 1.02. I-43

¹H NMR (400 MHz, DMSO): δ 9.42 (s, 1H), 8.00 (app q, J = 8.2 Hz, 2H),7.52 (q, J = 8.1 Hz, 1H), 7.30-7.26 (m, 2H), 7.15 (t, J = 7.5 Hz, 1H),4.30 (br s, 4H), 3.87 (s, 3H), 2.52 (s, 3H). MS (m/z): 289.1 (M + H)⁺;r.t. = 1.372.

Methyl 3-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-4-fluorobenzoate(I-44)

Methyl 3-amino-4-fluorobenzoate (8.4 mmol) and tert-butyl(2-oxoethyl)carbamate (12.6 mmol) were dissolved in MeOH (100 mL) andAcOH (10 mL). After stirring for 1 hr at room temperature the reactionmixture was treated with NaCNBH₃ (16.6 mmol) and stirred for 2 hrs.After removal of solvent (aspirator) the mixture was dissolved in EtOAcand washed with NaHCO₃ (sat., 1×). The aqueous phase was washed withEtOAc (1×) and the combined organic layers were dried (MgSO₄). Theproduct (I-44) was taken on crude for the next step. ¹H NMR (400 MHz,CDCl3): δ 7.40-7.38 (m, 2H), 7.02 (dd, J=8.8 Hz, 11.1, 1 H), 4.88 (s,1H), 3.91 (s, 3H), 3.40-3.38 (m, 4H), 1.47 (s, 9H). MS (m/z): 313.1(M+H)⁺.

Methyl 3-((2-aminoethyl)amino)-4-fluorobenzoate (I-45)

Methyl 3-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-4-fluorobenzoate(1.57 g) was dissolved in dichloromethane (10 mL) and treated with TFA(10 mL). The reaction stirred at room temperature for 1 hr. The solventwas azeotropically removed with dichloromethane and the cream coloredsolid (I-45) was used directly for the next step. ¹H NMR (400 MHz,DMSO): δ 7.78 (s, 3H), 7.24 (m, 3H), 5.93 (s, 1H), 3.83 (s, 3H),3.50-3.22 (m, 5H), 3.04 (s, 2H). MS (m/z): 213.1 (M+H)⁺.

Methyl 4-fluoro-3-((2-((3-fluorobenzyl)amino)ethyl)amino)benzoate (I-46)

A mixture of methyl 3-((2-aminoethyl)amino)-4-fluorobenzoate (5.0 mmol),sodium bicarbonate (5.0 mmol) and methanol (30 mL) stirred at roomtemperature for 30 mins. 3-fluoro-benzaldehyde (5.0 mmol) was added andstirred at room temperature for 2 hrs. Sodium Borohydride (7.4 mmol) wasadded in portions. The reaction stirred at room temperature for 30 mins.LCMS showed that the reaction was complete. The solvent was removed andthe residue was diluted with ethyl acetate (50 mL). The organic solutionwas washed with water and brine, dried over MgSO₄, and concentrated. Thecrude material was purified by flash chromatography (silica 5-40% ethylacetate/hexanes) to give (I-46) as a white solid. ¹H NMR (400 MHz,DMSO): δ 7.33 (td, J=6.3, 8.1 Hz, 1H), 7.25 (dd, J=1.9, 8.6 Hz, 1H),7.23-7.10 (m, 4H), 7.08-6.98 (m, 1H), 5.67 (s, 1H), 3.82 (s, 3H), 3.74(s, 2H), 3.20 (q, J=6.2 Hz, 2H), 2.72 (t, J=6.3 Hz, 2H). MS (m/z): 321.1(M+H)⁺.

The following intermediates were prepared according to the proceduresdescribed for the synthesis of I-46 using the appropriate reagents.

Compound Structure Characterization Data I-47

MS (m/z): 317.1 (M + H)⁺; r.t. = 1.274 I-48

¹H NMR (400 MHz, CDCl₃): δ 7.38-7.31 (m, 3H), 7.27-7.21 (m, 1H), 7.11(1H, td, J = 1.1, 7.4 Hz, 1H), 7.07-6.96 (m, 2H), 4.50 (m, 1H),3.91-3.85 (m, 5H), 3.34-3.24 (m, 2H), 2.98-2.90 (m, 2H). I-49

¹H NMR (400 MHz, DMSO): δ 8.88 (br s, 1H), 7.78-7.66 (m, 3H), 7.60 (d, J= 8.0 Hz, 2H), 7.34-7.22 (m, 3H), 5.02 (br s, 1H), 4.32 (t, J = 4.9 Hz,2H), 3.91 (s, 3H), 3.63 (t, J = 4.9 Hz, 2H), 3.12 (br s, 2H). MS (m/z):303.1 (M + H)⁺; r.t. = 1.71.

Methyl 4-(2-aminoethoxy)-3-fluorobenzoate (I-51)

To a mixture of methyl 3-fluoro-4-hydroxybenzoate (17 mmol), cesiumcarbonate (25 mmol) and DMF (30 mL) was added tert-butyl(2-bromoethyl)carbamate (20 mmol). The resulting suspension stirred at60° C. for 4 hrs. The reaction was diluted with water (100 mL) and theresulting precipitate was collected by vacuum filtration. The filtercake was washed with water and dried under vacuum to give pure I-50.I-50 (17 mmol) was taken up in dichloromethane (12 mL) and treated withTFA (5 mL). The reaction stirred at room temperature for 30 mins. LCMSshowed that the reaction was complete. The reaction was concentrated todryness. The crude material was taken up in dichloromethane and stirredwith solid sodium bicarbonate for 8 hrs to remove any residual TFA. Themixture was filtered and the filtrate was dried to give I-51 as a clearoil. ¹H NMR (400 MHz, DMSO): δ 7.79 (d, J=9.4 Hz, 1H), 7.73 (dd, J=1.6,11.8 Hz, 1H), 7.33 (t, J=8.6 Hz, 1H), 4.22 (t, J=5.4 Hz, 2H), 3.84 (s,3H), 3.12 (t, J=5.4 Hz, 2H).

Methyl 4-(2-(benzylamino)ethoxy)-3-fluorobenzoate (I-52)

A mixture of methyl 4-(2-aminoethoxy)-3-fluorobenzoate (I-51) (1.4 mmol)and sodium bicarbonate (1.4 mmol) in methanol (5 mL) was stirred at roomtemperature for 30 mins. Acetic acid (1.0 mL) and aldehyde was added andthe reaction stirred at room temperature for 2 hrs. NaCNBH₃ (2.1 mmol)was added in three portions. The reaction stirred at room temperaturefor an additional 15 mins. LCMS indicated that the reaction wascomplete. The reaction was diluted with water and extracted with ethylacetate. The organic extracts were combined and washed with water andbrine, dried over MgSO₄ and concentrated to give I-52. No furtherpurification was necessary. MS (m/z): 304.1 (M+H)⁺.

The following intermediate was prepared according to the proceduredescribed for the synthesis of I-52 using the appropriate reagents.

Compound Structure Characterization Data I-53

MS (m/z): 322.1 (M + H)⁺; r.t. = 1.31

Methyl4-(2-((tert-butoxycarbonyl)(2-fluorobenzyl)amino)ethoxy)-3-fluorobenzoate(I-55)

Di-tert-butyl dicarbonate (3.0 mmol) was added to a mixture of2-((2-fluorobenzyl)amino)ethanol (3.0 mmol) in dichloromethane (25 mL)and NaOH (1 M in H₂O, 9.6 mL, 9.6 mmol) at room temperature. Thereaction mixture was stirred for 16 hrs and then diluted with H₂O (50mL) and dichloromethane (25 mL). Upon separation of the layers, theaqueous phase was washed with dichloromethane (2×, 50 mL) and thecombined organic extracts were washed with H₂O (1×50 mL), dried (Na₂SO₄)and concentrated (aspirator). The crude mixture was purified bychromatography (silica gel, 0-60% EtOAc/hexanes) to give I-54 as an oil.Diisopropyl azodicarboxylate (0.41 mL, 2.1 mmol) was added dropwise to amixture of I-54 (1.0 mmol), methyl 3-fluoro-4-hydroxybenzoate (2.0 mmol)and PPh₃ (2.0 mmol) in THF (15 mL) at 0° C. After stirring for 10 mins,the reaction mixture was allowed to warm to room temperature on its ownaccord and stirred for an additional 4 hrs. The reaction mixture wasdiluted with Et₂O (50 mL) and H₂O (50 mL) and the layers were separated.The aqueous phase was washed with Et₂O (2×, 50 mL) and the combinedorganic extracts were dried (MgSO₄) and then concentrated (aspirator).The crude mixture was purified by chromatography (silica gel, 0-25%EtOAc/hexanes) to give an inseparable mixture of I-55 and methyl3-fluoro-4-hydroxybenzoate. The mixture was concentrated, diluted withEtOAc (100 mL) and then washed with NaOH (×2, 1 M, 25 mL) to removemethyl 3-fluoro-4-hydroxybenzoate from the mixture. The organic phasewas dried (MgSO₄) and concentrated (aspirator) to give I-55. ¹H NMR (400MHz, CDCl₃): δ 7.69-7.55 (m, 2H), 7.23-7.06 (m, 2H), 7.00-6.86 (m, 2H),6.86-6.71 (m, 1H), 4.56-4.45 (m, 2H), 4.14-3.99 (m, 2H), 3.75 (s, 3H),3.59-3.44 (m, 2H), 1.40-1.25 (m, 9H).

Methyl 3-fluoro-4-(2-((2-fluorobenzyl)amino)ethoxy)benzoate (I-56)

Trifluoroacetic acid (2 mL) was added to a solution of I-55 (0.9 mmol)in dichloromethane (8 mL) at room temperature. After stirring for 1 hrthe reaction mixture was concentrated (aspirator), diluted with MeOH andneutralized by passing the mixture through SPE-carbonate polymer boundcartridges (6 cartridges, 100 mg units each). The solvent was removed(aspirator) to give I-56. MS (m/z): 322.1 (M+H)⁺; r.t.=0.981.

Methyl 3-(2-aminoacetamido)-4-fluorobenzoate (I-58)

HATU (8.2 mmol) was added to a mixture of N-Boc glycine (6.8 mmol),methyl 3-amino-4-fluorobenzoate (6.2 mmol), diisopropylethylamine (3.6mL, 20.5 mmol) and DMF (15 mL). The reaction was stirred at roomtemperature for 1 hr. LCMS indicated that the reaction was complete. Thereaction was diluted with water and extracted with ethyl acetate. Theorganic extracts were dried over MgSO₄ and concentrated. No furtherpurification was necessary. The crude amide (I-57) (9.2 mmol) wasdissolved in dichloromethane (15 mL) and treated with trifluoroaceticacid (8 mL). The reaction stirred at room temperature for 1 hr. LCMSindicated that the reaction was complete. The reaction was concentratedto dryness and the crude material was purified by preparative HPLC togive I-58. ¹H NMR (400 MHz, d₄-MeOH) δ 8.71 (br d, J=8.0 Hz, 1H),7.79-7.75 (m, 1H), 7.21 (dd, J=10.3, 8.2 Hz, 1H), 3.87 (s, 2H), 3.82 (s,3H). MS (m/z): 227.1 (M+H)⁺.

Methyl 4-fluoro-3-(2-((3-methylbenzyl)amino)acetamido)benzoate (I-59)

A mixture of methyl 3-(2-aminoacetamido)-4-fluorobenzoate (I-58) (1.5mmol) and sodium bicarbonate (1.5 mmol) in methanol (5 mL) was stirredat room temperature for 30 mins. Acetic acid (1.0 mL) and3-methylbenzaldehyde were added and the reaction stirred at roomtemperature for 2 hrs. NaCNBH₃ (2.2 mmol) was added in three portions.The reaction stirred at room temperature for an additional 15 mins. LCMSindicated that the reaction was complete. The reaction was concentratedand purified by preparative HPLC to give I-59. MS (m/z): 331.1 (M+H)⁺;r.t.=1.51.

The following intermediate was prepared according to the proceduredescribed for the synthesis of I-59 using the appropriate reagents.

Compound Structure Characterization Data I-59A

¹H NMR (400 MHz, CDCl₃): δ 9.00 (s, 1H), 8.11 (s, 1H), 8.00 (d, J = 8.1,1H), 7.81 (d, J = 7.8, 1H), 7.43 (t, J = 7.9, 1H), 7.31 (m, 2H),7.19-7.12 (m, 3H), 4.41-4.38 (m, 2H), 3.92 (s, 3H), 3.47 (s, 2H).

(S)-Methyl3-(2-((tert-butoxycarbonyl)amino)propanamido)-4-fluorobenzoate (I-60)

Iso-Butyl chloroformate (5.3 mmol) was added to a solution ofN-(tert-butoxycarbonyl)-L-alanine (5.0 mmol) and N-methylmorpholine (5.3mmol) in THF (25 mL) at 0° C. After stirring for 30 min, methyl3-amino-4-fluorobenzoate (5.3 mmol) was added as a solid and theresulting mixture was stirred for 16 hrs at room temperature. Afterremoval of the solvent (aspirator), the residue was dissolved in EtOAc(50 mL) and washed with NaHCO₃ (sat., 50 mL), HCl (0.1 M in H₂O, 50 mL)and brine (50 mL). The organics was dried (MgSO₄) and concentrated(aspirator). The crude material was purified by chromatography (silicagel, 0-40% EtOAc/hexanes) to give I-60. ¹H NMR (600 MHz, CDCl₃): δ 8.96(dd, J=2.0, 7.6 Hz, 1H), 8.67 (br s, 1H), 7.83-7.75 (m, 1H), 7.17-7.11(m, 1H), 4.94 (br s, 1H), 4.36 (br s, 1H), 3.90 (s, 3H), 1.50-1.43 (m,12H).

(S)-Methyl 4-fluoro-3-(2-((2-fluorobenzyl)amino)propanamido)benzoate(I-62)

Trifluoroacetic acid (1.0 mL) was added to a solution of I-60 (0.9 mmol)in dichloromethane (5 mL) at room temperature. After stirring for 30mins, the solvent was removed (aspirator) and the residue was dissolvedin dichloromethane (25 mL) and washed with NaHCO₃ (sat., 25 mL). Theaqueous phase was washed with dichloromethane (2×, 25 mL) and thecombined organic extracts were dried (Na₂SO₄) and concentrated(aspirator) to give 190 mg of I-61 as an oil. 2-Fluorobenzaldehyde (94mg, 0.8 mmol) was added to a solution of I-61 (0.8 mmol) in MeOH (9 mL)and AcOH (1 mL). After stirring for 1 hr at room temperature, thereaction mixture was treated with NaBH₄ (1.6 mmol) and stirred for 30mins. After removal of solvent (aspirator), the mixture was dissolved inEtOAc (25 mL) and washed with NaHCO₃ (sat., 25 mL). The aqueous phasewas washed with EtOAc (2×, 25 mL) and the combined organic extracts weredried (MgSO₄) and concentrated (aspirator). The material was purified bychromatography (silica gel, 0-60% EtOAc/hexanes) to give I-62. MS (m/z):349.2 (M+H)⁺; r.t.=1.097.

1-methyl-4,5-dihydro-1H-benzo[2,3]oxepino[4,5-c]pyrazole-3-carboxylicacid (I-64)

Intermediate I-64 was prepared according to the procedure described forthe preparation of intermediate I-4, starting with3,4-dihydrobenzo[b]oxepin-5(2H)-one (I-63, commercially available). ¹HNMR (400 MHz, DMSO): ¹H NMR (400 MHz, DMSO): δ 12.77 (s, 1H), 7.80 (d,J=2.6 Hz, 1H), 7.44 (dd, J=8.6, 2.6 Hz, 1H), 7.22 (d, J=8.7 Hz, 1H),4.33 (t, J=6.3 Hz, 2H), 4.05 (s, 3H), 3.12 (t, J=6.3 Hz, 2H). MS (m/z):279.0/281.0 (M+H)⁺ (chlorine isotope pattern); r.t.=1.302.

2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)aceticacid (I-66)

Step 1

To a 20 mL reaction vessel was added reagents in the following order:glycine (10 mmol), AcOH (2 mL), MeOH (12 mL), water (2 mL). This wasstirred until complete dissolution. Next was added 2-fluorobenzaldehyde(5.0 mmol). After 20 mins, sodium cyanoborohydride (3.0 mmol) was added.The reaction was stirred 10 mins, at which time an additional portion ofsodium cyanoborohydride was added (3.0 mmol). The reaction was stirredfor 10 mins, filtered, and then purified by acetic acid modified (0.05%)reverse phase-chromatography (10 to 50%, water-ACN) and subsequentlyrecrystallized by MeOH/water 1:5 (2 mL) to furnish2-((2-fluorobenzyl)amino)acetic acid (I-65) as a white powder. ¹H NMR(400 MHz, DMSO): δ 7.61 (t, J=8.1 Hz, 1H), 7.54 (app q, J=7.8 Hz, 1H),7.36-7.28 (m, 2H), 4.24 (s, 2H), 3.92 (s, 2H). MS (m/z): 184.1 (M+H)⁺.

Step 2

To a 100 mL reaction vessel was added reagents in the following order:8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-4) (1.5 mmol), THF (15 mL), and N-methyl morpholine (NMM) (1 mL, 6.9mmol). This was stirred until complete dissolution. Next was added2-chloro-4,6-dimethoxy-1,3,5-triazine (1.6 mmol) and this solution wasstirred for 40 mins at 50° C. until a white precipitate fully formed.The precipitate was physically agitated with rapid stirring to ensureall solids were well mixed. Next was added2-((2-fluorobenzyl)amino)acetic acid (I-65) (2.3 mmol) and the reactionwas stirred for 30 mins at 50° C. and then diluted with 5 mL of MeOH andthen purified by acetic acid modified (0.05%) reverse phasechromatography (30 to 80%, water/ACN) and subsequently recrystallized byMeOH/water 1:1 (20 mL) to furnish a white powder (I-66). ¹H NMR (400MHz, CDCl₃): δ 10.05 (br s, 1H), 7.39 (app t, J=7.8 Hz, 2H), 7.23-7.20(m, 1H), 7.14 (d, J=8.3 Hz, 1H), 7.09 (t, J=8.2 Hz, 1H), 7.02 (t, J=8.2Hz, 1H), 6.84 (d, J=8.3 Hz, 2H), 5.44-5.40 (m, 2H), 4.82 (br s, 1H),4.59 (br s, 1H), 4.05-4.02 (m, 1H), 4.01 (s, 3H). MS (m/z): 430.1/432.1(M+H)⁺ (chlorine isotope pattern).

2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)aceticacid (I-68)

Step 1

To a suspension of the8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-4) (1.9 mmol) in CH₂Cl₂ (10 mL) was added catalytic amount of DMF (25uL) and dropwise oxalyl chloride (0.78 mL, 8.9 mmol) at 0° C. Theresulting suspension was warmed up to room temperature and stirred for 1hr. The solvent was removed under vacuum to dryness completely (need toremove oxalyl chloride completely). The obtained residue was dissolvedwith CH₂Cl₂ (10 mL) and then dropwise into a solution of methyl2-((3-fluorobenzyl)amino)acetate (1.9 mmol) in CH₂Cl₂ (10 mL) in thepresence of DIEA (0.56 mL, 3.8 mmol). The reaction mixture was stirredfor 30 min at room temperature and then was charged 50 mL water. Organiclayer was separated and aqueous layer was extracted with CH₂Cl₂ (50 mL).Combined organic layers were washed with H₂O and brine successively,dried over Na₂SO₄, filtered and concentrated to give the crude product,which was purified by column chromatography (0-60% EtOAc in hexanes) togive methyl2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetate(I-67). ¹H NMR (400 MHz, CDCl₃): δ 7.47-7.43 (m, 2H), 7.34-7.29 (m, 1H),7.23-7.19 (m, 1H), 7.18-7.04 (m, 2H), 6.97 (dd, J=2.3, 8.7 Hz, 1H),5.57-4.12 (m, 9H), 3.77-3.74 (m, 3H). Mixture of rotamers. MS (m/z):444.1 (M+H)⁺.

Step 2

Methyl2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetate(I-67) (1.6 mmol) was dissolved in THF/MeOH/H₂O (3:2:1, 10 mL) andfollowed by addition of LiOH monohydrate (0.408 g, 9.72 mmol). Thereaction mixture was stirred at room temperature for 1 hr and dilutedwith 10 ml of water and acidified to pH=2.0. Solid was collected anddried to yield2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)aceticacid (I-68). ¹H NMR (400 MHz, DMSO): δ 12.73 (s, 1H), 7.71 (t, J=2.6 Hz,1H), 7.42-7.30 (m, 3H), 7.24-7.15 (m, 2H), 7.07-7.02 (m, 1H), 5.41-4.10(m, 9H). Mixture of rotamers. MS (m/z): 430.0 (M+H)⁺.

Methyl 3-(2-((cyclopentylmethyl)amino)acetamido)-4-fluorobenzoate (I-70)

Step 1

To a 40 mL reaction vessel was added reagents in the following order:N-(tert-butoxycarbonyl)glycine (1.0 mmol), THF (4.0 mL), N-methylmorpholine (c.a. 0.2 mL, 2.0 mmol). This was stirred until completedissolution. Next was added 2-chloro-4,6-dimethoxy-1,3,5-triazine (1.0mmol) and this solution was stirred for 20 mins at 50° C. until a whiteprecipitate fully formed. The precipitate was physically agitated toensure all solids were well stirred. Next was added methyl3-amino-4-fluorobenzoate (1.3 mmol) and the reaction was stirred for 30mins at 50° C. and then at room temperature for 2 hrs, then diluted with5 mL of MeOH and then purified by being poured into water (rapidlystirred, 25 mL). A white solid was collected and treated withdichloromethane (5 mL) and TFA (5 mL), and heated to 34° C. for about 1hr until removal of the Boc group was complete (by LCMS monitoring). Thematerial was concentrated to dryness and subjected to reverse-phasechromatography (10 to 40% water/ACN) to give I-69. The desired fractionswere then passed through polymer bound ion-exchange free base cartridgesto removal any residual TFA (using product StratoSpheres, SPE PL HCO3 MPSPE 0.9 mmol nominal load×2 units). Mobilizing eluent was MeOH (10 mLtotal flush volume). ¹H NMR (400 MHz, d₄-MeOH): δ 8.71 (br d, J=8.0 Hz,1H), 7.79-7.75 (m, 1H), 7.21 (dd, J=10.3, 8.2 Hz, 1H), 3.87 (s, 2H),3.82 (s, 3H). MS (m/z): 227.1 (M+H)⁺.

Step 2

To a 20 mL reaction vessel was added reagents in the following order:methyl 3-(2-aminoacetamido)-4-fluorobenzoate (I-69) (0.63 mmol,), AcOH(1 mL), MeOH (4 mL) and cyclopentanecarbaldehyde (1 mmol). After 30mins, sodium cyanoborohydride (0.80 mmol) was added. The reaction wasstirred 10 mins, at which time an additional portion of sodiumcyanoborohydride was added (0.40 mmol). The reaction was stirred for 2hrs, filtered, and then purified TFA modified (0.05%) reversephase-chromatography (20 to 60%, water-ACN) and subsequently residualTFA was removed using a polymer bound ion exchange cartridge (SPE PLHCO₃ MP SPE 0.9 mmol nominal load x 1 units and 8 mL MeOH as mobilizingeluent) to give the amine (I-70) as a glassy solid. ¹H NMR (400 MHz,d₄-MeOH): δ 8.80 (br d, J=8.0 Hz, 1H), 7.91-7.86 (m, 1H), 7.32 (dd,J=11.0, 8.0 Hz, 1H), 4.08 (s, 2H), 3.92 (s, 3H), 3.08 (app d, J=8.1 Hz,2H), 2.31-2.22 (m, 1H), 1.97-1.92 (m, 2H), 1.77-1.64 (m, 4H), 1.36-1.30(m, 2H). MS (m/z): 309.1 (M+H)⁺.

The following intermediates were prepared according to the proceduresdescribed for the synthesis of I-70 using the appropriate reagents.

Compound Structure Characterization Data I-71

MS (m/z): 291.1 (M + H)⁺; r.t. = 1.238

8-chloro-N-(2-fluorobenzyl)-N-(2-hydroxyethyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamide(I-73)

Step 1

To a 250 mL reaction vessel was added reagents in the following order:ethanol amine (40.0 mmol,), AcOH (8 mL), MeOH (60 mL), and2-fluorobenzaldehyde (20.0 mmol). After 30 mins, sodium cyanoborohydride(20.0 mmol) was added portionwise over 30 mins. The reaction was stirred1 hr, and then partially concentrated (in vacuo) before directpurification by acetic acid modified (0.05%) reversephase-chromatography (5 to 15%, water-ACN) to furnish upon drying2-((2-fluorobenzyl)amino)ethanol (I-72) as a semi-solid. ¹H NMR (400MHz, d₄-MeOH): δ 7.61 (dt, J=7.9, 1.8 Hz, 1H), 7.41-7.35 (m, 1H), 7.21(app t, J=7.9 Hz, 1H), 7.13 (app t, J=8.9 Hz, 1H), 4.00 (s, 2H), 3.73(t, J=6.2 Hz, 1H), 2.87 (t, J=6.2 Hz, 1H). MS (m/z): 170.1 (M+H)⁺.

Step 2

To a 100 mL reaction vessel was added reagents in the following order:8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-4) (2.0 mmol, THF (10 mL) and N-methyl morpholine (0.72 mL, 5.0mmol). This was stirred until complete dissolution. Next was added2-chloro-4,6-dimethoxy-1,3,5-triazine (2.2 mmol) and this solution wasstirred for 20 mins at 50 C until a white precipitate fully formed. Theprecipitate was physically agitated with rapid stirring to ensure allsolids were well mixed. Next was added 2-((2-fluorobenzyl)amino)ethanol(I-72) (2.4 mmol) and the reaction was stirred for 30 mins at 50° C. andthen diluted with 5 mL of MeOH and then purified by acetic acid modified(0.05%) reverse phase chromatography (30 to 80%, water/ACN) andsubsequently recrystallized by MeOH/water 1:3 (10 mL) to furnish a whitepowder (I-73). ¹H NMR (400 MHz, DMSO): δ 7.74-7.70 (m, 1H), 7.38-7.28(m, 5H), 7.04 (dd, J=8.4, 1.5 Hz, 1H), 5.40-5.35 (m, 3H), 4.80-4.75 (m,2H), 4.20 (s, 3H), 3.93 (t, J=6.2 Hz, 1H) 3.63 (q, J=6.2 Hz, 1H), 3.55(q, J=6.2 Hz, 1H), 3.41 (t, J=6.5 Hz, 1H). Mixture of rotamers. MS(m/z): 416.1/418.1 (M+H)⁺ (chlorine isotope pattern).

The following intermediates were prepared according to the proceduresdescribed for the synthesis of I-73 using the appropriate reagents.

Compound Structure Characterization Data I-74

¹H NMR (400 MHz, DMSO): δ 7.78-7.75 (m, 2H), 7.30-7.16 (m, 5H),5.40-5.35 (m, 2H), 4.80 (br s, 2H), 4.04 (s, 3H), 3.78 (app t, J = 5.0Hz, 2H), 3.34 (app t, J = 5.0 Hz, 2H), 3.45 (t, J = 4.0 Hz, 1H). Mixtureof rotamers. MS (m/z): 398.1 (M + H)⁺. I-75

¹H NMR (400 MHz, DMSO): δ 7.73-7.70 (m, 2H), 7.36-7.00 (m, 5H),5.35-5.30 (m, 2H), 4.78 (br s, 2H), 4.14 (s, 3H), 3.88 (app t, J = 5.0Hz, 2H), 3.45-3.50 (m, 2H), 3.38-3.36 (m, 1H). Mixture of rotamers. MS(m/z): 400.1 (M + H)⁺. I-76

¹H NMR (400 MHz, DMSO): δ 7.75-7.69 (m, 2H), 7.46-6.92 (m, 6H),5.25-5.15 (m, 2H), 4.73 (br s, 2H), 4.04 (s, 3H), 3.88-3.24 (m, 4H).Mixture of rotamers. MS (m/z): 382.1 (M + H)⁺. I-77

MS (m/z): 390.1 (M + H)⁺; r.t. = 1.289

1,5,5-trimethyl-4,5-dihydro-1H-benzo[g]indazole-3-carboxylic acid (I-78)

Intermediate I-78 was prepared according to the procedure described forthe preparation of intermediate I-4, starting with4,4-dimethyl-3,4-dihydronaphthalen-1(2H)-one. MS (m/z): 257.1 (M+H)⁺.

Example 1

4-Fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid (Compound 1) Step 1

To a solution of methyl4-fluoro-3-(2-((2-fluorobenzyl)amino)acetamido)benzoate (I-17) (8.2mmol) and HATU (8.2 mmol) in DMF (30 mL), diisopropylethylamine (30mmol) and8-fluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-5) (7.4 mmol) were added to give a yellowish solution. The reactionmix was stirred at room temperature for 3 hr. The reaction mixture waspoured onto ice 300 mL resulting in a precipitate that was collected byvacuum filtration. The solid was then re-dissolved in EtOAc (600 mL) andwashed with 5% Na₂CO₃, water, brine, dried and concentrated The solidwas purified by trituration in hot MeOH. The resulting solid wasfiltered and dried. ¹H NMR (400 MHz, DMSO): δ 10.14 (m, 1H), 8.79-8.46(m, 1H), 7.80-7.66 (m, 1H), 7.56 (m, 1H), 7.50-7.29 (m, 3H), 7.22 (m,3H), 7.06 (m, 1H), 5.38 (m, 3H), 4.82 (d, J=41.7 Hz, 3H), 4.35-4.03 (m,3H), 3.85 (m, 3H). Mixture of rotamers.

Step 2

The resulting ester (5 mmol) was dissolved in a 4:1 THF-MeOH (100 mL)and treated with 1N KOH (20 mL). The white suspension was stirred atroom temperature for 2 hrs resulting in a clear solution. The reactionwas stirred at room temperature for an additional 16 hrs until thereaction was completed as indicated by LCMS. The organic solvent wasremoved in vacuo resulting in a white suspension. The resultingsuspension was diluted with water (100 mL) and the pH was adjusted to ˜5w/AcOH. The white solid was collected by filtration, washed with waterand dried under vacuum for 24 hrs. The carboxylic acid product wasfurther purified by stirring in 8:2 MeOH-water solution (200 mL) at 70°C. for 2 hrs. After cooling to room temperature, the solid was collectedby vacuum filtration to give4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid. ¹H NMR (400 MHz, DMSO): δ 10.06-10.01 (m, 1H), 8.58-8.56 (m, 1H),8.51-8.49 (m, 1H), 7.72-7.69 (m, 1H), 7.58-7.53 (m, 1H), 7.46-7.33 (m,3H), 7.24-7.12 (m, 3H), 7.08-7.04 (m, 1H), 5.40-5.37 (m, 3H), 4.86-4.76(m, 3H), 4.24-4.12 (m, 4H). Mixture of rotamers. MS (m/z): 551.1 (M+H)⁺.

In one embodiment, a solution of L-arginine (0.82 mmol) in deionizedwater (6 mL) was added to a suspension of4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3carboxamido)acetamido)benzoic acid (0.82 mmol) in MeOH (50 mL). Theclear mixture was stirred for 0.5 hr at room temperature and thenconcentrated to give a white semi-solid residue. The residue was takenin anhydrous acetonitrile (25 mL) and the white suspension was slowlyconcentrated under reduced pressure. The process was repeated severaltimes (5×) to give4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid L-arginine salt. Melting point (157° C.). ¹H NMR (400 MHz,DMSO): δ 9.88-9.77 (m, 1H), 8.37-8.26 (m, 1H), 8.03-7.68 (m, 5H),7.66-7.51 (m, 2H), 7.48-7.31 (m, 3H), 7.15 (m, 6H), 5.43-5.35 (m, 2H),4.86-4.72 (m, 3H), 4.22-4.10 (m, 4H), 3.26-3.18 (m, 1H), 3.17-2.99 (m,2H), 1.76-1.52 (m, 4H).

In another embodiment, suspension of4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid (0.84 mmol) in MeOH (20 mL) was heated to 70° C. A 1N solution ofNaOH (0.84 mmol) was added drop wise. The clear solution was stirred atroom temperature for 1 hr before the solvent was removed under reducedpressure. The crude material was triturated with MeOH and crystallizedby slurring in acetonitrile (5 mL) to give4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid sodium salt. Melting point (161° C.). ¹H NMR (400 MHz, DMSO): δ9.86-9.60 (m, 1H), 8.27-8.17 (m, 1H), 7.63-7.55 (m, 2H), 7.48-7.31 (m,2H), 7.26-7.04 (m, 5H), 5.42-5.37 (m, 2H), 4.84-4.74 (m, 3H), 4.20-4.12(m, 4H).

4-Fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid (Compound 2)

4-Fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid was prepared following the procedures for Compound 1. Melting point(246° C.). ¹H NMR (400 MHz, DMSO): δ 10.14-10.04 (m, 1H), 8.71-8.58 (m,1H), 7.79-7.69 (m, 1H), 7.62-7.51 (m, 1H), 7.47-7.36 (m, 2H), 7.29-7.02(m, 5H), 5.42-5.32 (m, 2H), 4.84-4.72 (m, 2H), 4.23-4.08 (m, 3H),3.88-3.80 (m, 3H). Mixture of rotamers. MS (m/z) (M+H)⁺, 551.3. Anal.Calcd for C₂₈H₂₁F₃N₄O₅*0.15 CH₄O: C, 61.09; H, 3.85; N, 10.18. Found: C,60.97; H, 4.02; N, 10.16.

Methanol (500 mL) and dichloromethane (500 mL) were added to a mixtureof4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid (22 mmol) and 2-amino-2-hydroxymethyl-propane-1,3-diol(TRIS; 22 mmol). The reaction was stirred at 25° C. for 3 hrs. Thesolvent was removed in vacuum, and dichloromethane (500 mL) was added tothe solid and concentrated (repeated 2×). The solid was suspended in 100mL of dichloromethane and collected by vacuum filtration to give4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid TRIS salt. Melting point (125° C.). ¹H NMR (400 MHz, DMSO)δ 10.01-9.65 (m, 1H), 8.59-8.30 (m, 1H), 7.76-6.95 (m, 10H), 5.53-4.00(m, 9H), 3.58-3.10 (m, 10H). Anal. Calcd for C₃₂H₃₂F₃N₅O₈*1 H₂O: C,55.73; H, 4.97; N, 10.16. Found: C, 55.74; H, 4.99; N, 10.12.

In one embodiment, a solution of L-arginine (0.73 mmol) in deionizedwater (5.8 mL) was added to suspension of4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid (0.73 mmol) in methanol (70 mL). The mixture was heated to 70° C.for 0.5 hrs. After cooling to room temperature, the solvent was removedin vacuum. A 2:1 mixture of acetonitrile and methanol (20 mL) was addedto the solid and concentrated (repeated 2×). The solid was crystallizedby slurring the solid residue in a 2:1 mixture of acetonitrile andmethanol (5 mL) and stirring the suspension at room temperature for 24hrs to give4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid L-arginine salt as a white precipitate. Melting point (206°C.). ¹H NMR (400 MHz, DMSO) δ 9.88-9.78 (m, 1H), 8.41-8.31 (m, 1H),8.08-7.82 (m, 5H), 7.68-7.52 (m, 2H), 7.47-7.36 (m, 1H), 7.28-7.04 (m,6H), 5.43-5.30 (m, 3H), 4.81-4.70 (m, 3H), 4.14 (s, 4H), 3.26-3.21 (m,2H), 3.16-3.01 (m, 2H), 1.78-1.52 (m, 4H).

In another embodiment, a solution of L-lysine (0.20 mmol) in deionizedwater (3 mL) was added to a hot suspension of4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid (0.20 mmol) in methanol (25 mL). The mixture was stirred at roomtemperature for 16 hrs. The solvent was removed under vacuum and thecrude material was crystallized by slurring the solid residue inacetonitrile (5 mL). The mixture was stirred at room temperature foradditional 24 hrs to give4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoic acid L-lysine salt as a white precipitate. ¹H NMR (400 MHz,DMSO): δ 9.86-9.77 (m, 1H), 8.40-8.28 (m, 1H), 7.68-7.51 (m, 2H),7.48-7.36 (m, 1H), 7.29-7.02 (m, 7H), 5.43-5.36 (m, 2H), 5.36-5.31 (m,1H), 4.80-4.71 (m, 3H), 4.19-4.11 (m, 4H), 3.17-3.09 (m, 1H), 2.76-2.64(m, 2H), 1.77-1.31 (m, 7H); some peaks fall under solvent peaks.

The following compounds were prepared according to the proceduresdescribed in Example 1 using the appropriate intermediates.

Compound Structure Characterization Data  3

¹H NMR (400 MHz, DMSO): δ 10.03 (s, 1H), 8.72-8.39 (m, 1H), 7.81-7.59(m, 2H), 7.48-6.86 (m, 7H), 5.54-5.19 (m, 3H), 4.79-4.61 (m, 2H),4.22-4.03 (m, 4H), 2.30 (s, 3H). Mixture of rotamers. MS (m/z):563.2/565.2 (M + H)⁺ (chlorine isotope pattern).  4

¹H NMR (400 MHz, DMSO): δ 10.06-9.86 (m, 1H), 8.60-8.45 (m, 1H),7.80-7.61 (m, 2H), 7.42-6.92 (m, 8H), 5.48-5.22 (m, 3H), 4.77-4.66 (m,2H), 4.20-4.02 (m, 4H), 2.30 (s, 3H). Mixture of rotamers. MS (m/z):529.2 (M + H)⁺.  5

¹H NMR (400 MHz, DMSO): δ 10.31- 10.18 (m, 1H), 8.28-8.16 (m, 1H), 7.83-7.74 (m, 1H), 7.65-7.58 (m, 1H), 7.56- 7.31 (m, 4H), 7.26-7.17 (m, 2H),7.13- 7.06 (m, 1H), 6.96-6.85 (m, 1H), 5.48- 5.25 (m, 3H), 4.88-4.70 (m,2H), 4.21- 4.00 (m, 4H), 2.34-2.25 (m, 3H). Mixture of rotamers. MS(m/z): 529.2 (M + H)⁺.  6

¹H NMR (400 MHz, DMSO): δ 10.31- 10.21 (m, 1H), 8.29-8.17 (m, 1H), 7.82-7.74 (m, 1H), 7.66-7.49 (m, 2H), 7.49- 7.29 (m, 3H), 7.26-7.13 (m, 3H),7.02- 6.92 (m, 1H), 5.47-5.35 (m, 3H), 4.88- 4.71 (m, 3H), 4.21-4.02 (m,4H), 2.25- 2.14 (m, 3H). Mixture of rotamers. MS (m/z): 529.2 (M + H)⁺. 7

¹H NMR (400 MHz, DMSO): δ 10.31- 10.19 (m, J = 6.6 Hz, 1H), 8.26-8.16(m, J = 9.7 Hz, 1H), 7.83-7.74 (m, J = 2.9, 5.1 Hz, 1H), 7.65-7.49 (m,2H), 7.49-7.30 (m, 3H), 7.26-7.01 (m, 4H), 5.46-5.33 (m, J = 11.7 Hz,3H), 4.85-4.73 (m, J = 26.4 Hz, 2H), 4.20-4.04 (m, J = 10.6 Hz, 22.1,4H). Mixture of rotamers. MS (m/z): 533.1 (M + H)⁺.  8

¹H NMR (400 MHz, DMSO): δ 10.32- 10.21 (m, 1H), 8.29-8.17 (m, 1H), 7.83-7.68 (m, 15.0, 2H), 7.65-7.59 (m, 1H), 7.47-7.30 (m, 3H), 7.24-7.16 (m,2H), 6.98-6.87 (m, 2H), 5.49-5.38 (m, 3H), 4.88-4.69 (m, 2H), 4.18-4.02(m, 4H). Mixture of rotamers. MS (m/z): 533.2 (M + H)⁺.  9

¹H NMR (400 MHz, DMSO): δ 10.31- 10.20 (m, 1H), 8.27-8.17 (m, 1H), 7.82-7.58 (m, 3H), 7.48-7.30 (m, 3H), 7.26- 7.08 (m, 4H), 5.48-5.38 (m, 3H),4.85- 4.72 (m, 2H), 4.19-4.02 (m, 4H). Mixture of rotamers. MS (m/z):549.1 (M + H)⁺. 10

¹H NMR (400 MHz, DMSO): δ 10.30- 10.20 (m, 1H), 8.26-8.17 (m, 1H), 7.86-7.72 (m, 2H), 7.66-7.58 (m, 1H), 7.48- 7.30 (m, 3H), 7.27-7.16 (m, 3H),5.45- 5.38 (m, 3H), 4.85-4.72 (m, 2H), 4.18- 4.05 (m, 4H). Mixture ofrotamers. MS (m/z): 551.1 (M + H)⁺. 11

¹H NMR (400 MHz, DMSO): δ 10.29- 10.21 (m, 1H), 8.28-8.15 (m, 1H), 7.82-7.74 (m, 1H), 7.66-7.58 (m, 1H), 7.49- 7.30 (m, 5H), 7.26-7.16 (m, 2H),5.49- 5.38 (m, 3H), 4.87-4.73 (m, 2H), 4.19- 4.06 (m, 4H). Mixture ofrotamers. MS (m/z): 551.2 (M + H)⁺. 12

¹H NMR (400 MHz, DMSO): δ 10.32- 10.19 (m, 1H), 8.26-8.18 (m, 1H), 7.83-7.57 (m, 3H), 7.48-7.16 (m, 6H), 7.13- 6.96 (m, 2H), 5.47-5.35 (m, 3H),4.86- 4.72 (m, 2H), 4.18-4.04 (m, 4H). Mixture of rotamers. MS (m/z):515.1 (M + H)⁺. 13

¹H NMR (400 MHz, DMSO): δ 10.23 (m, 1H), 8.23-8.14 (m, 1H), 7.82-7.72(m, 1H), 7.70-7.57 (m, 2H), 7.47-7.30 (m, 3H), 7.25-7.16 (m, 2H),7.08-7.02 (m, 1H), 5.44-5.33 (m, 3H), 4.84-4.72 (m, 2H), 4.18-4.04 (m,4H), 2.32-2.25 (m, 3H). Mixture of rotamers. MS (m/z): 563.2 (M + H)⁺.14

¹H NMR (400 MHz, DMSO): δ 10.29- 10.19 (m, 1H), 8.25-8.17 (m, 1H), 7.82-7.73 (m, 1H), 7.64-7.48 (m, 2H), 7.48- 7.30 (m, 3H), 7.30-7.15 (m, 3H),5.45- 5.38 (m, 3H), 4.84-4.72 (m, 2H), 4.17- 4.05 (m, 4H), 2.21-2.14 (m,3H). Mixture of rotamers. MS (m/z): 563.2 (M + H)⁺. 15

¹H NMR (400 MHz, DMSO): δ 10.31- 10.21 (m, 1H), 8.27-8.17 (m, 1H), 7.82-7.73 (m, 1H), 7.67-7.47 (m, 3H), 7.47- 7.29 (m, 3H), 7.27-7.15 (m, 2H),5.53- 5.38 (m, 3H), 4.86-4.69 (m, 2H), 4.21- 4.07 (m, 4H). Mixture ofrotamers. MS (m/z): 567.1 (M + H)⁺. 16

¹H NMR (400 MHz, DMSO): δ 10.13- 10.00 (m, 1H), 8.65-8.50 (m, 1H), 7.76-7.65 (m, 2H), 7.46-7.15 (m, 6H), 7.12- 6.97 (m, 2H), 5.44-5.32 (m, 3H),4.91- 4.73 (m, 2H), 4.27-4.06 (m, 4H). Mixture of rotamers. MS (m/z):533.2 (M + H)⁺. 17

¹H NMR (400 MHz, DMSO): δ 10.13-9.97 (m, 1H), 8.65-8.49 (m, 1H),7.86-7.67 (m, 2H), 7.48-7.30 (m, 3H), 7.27-7.16 (m, 3H), 5.44-5.36 (m,3H), 4.87-4.73 (m, 2H), 4.27-4.07 (m, 4H). Mixture of rotamers. MS(m/z): 569.2 (M + H)⁺. 18

¹H NMR (400 MHz, DMSO): δ 10.11-9.98 (m, 1H), 8.69-8.47 (m, 1H),7.87-7.65 (m, 2H), 7.48-7.30 (m, 2H), 7.30-7.06 (m, 4H), 5.49-5.29 (m,3H), 4.83-4.70 (m, 2H), 4.22-4.05 (m, 4H). Mixture of rotamers. MS(m/z): 569.2 (M + H)⁺. 19

¹H NMR (400 MHz, DMSO): δ 10.09-9.96 (m, 1H), 8.67-8.52 (m, 1H),7.87-7.65 (m, 2H), 7.43-7.16 (m, 7H), 5.49-5.29 (m, 3H), 4.79-4.69 (m,2H), 4.20-4.06 (m, 4H). Mixture of rotamers. MS (m/z): 551.2 (M + H)⁺.20

¹H NMR (400 MHz, DMSO): δ 10.08-9.94 (m, 1H), 8.66-8.53 (m, 1H),7.76-7.66 (m, 1H), 7.61-7.50 (m, 1H), 7.44-7.31 (m, 1H), 7.30-7.21 (m,1H), 7.21-7.02 (m, 5H), 5.43-5.26 (m, 3H), 4.79-4.66 (m, 2H), 4.19-4.05(m, 4H), 2.31-2.27 (s, 3H). Mixture of rotamers. MS (m/z): 547.1 (M +H)⁺. 21

¹H NMR (400 MHz, DMSO): δ 10.03 (s, 1H), 8.66-8.42 (m, 2H), 7.78-7.65(m, 2H), 7.52-7.14 (m, 6H), 5.25-5.13 (m, 1H), 4.82-4.66 (m, 2H),4.29-4.20 (m, 4H), 3.01-2.83 (m, 4H). Mixture of rotamers. MS (m/z):532.2 (M + H)⁺. 22

¹H NMR (400 MHz, DMSO): δ 10.07-9.97 (m, 1H), 8.67-8.51 (m, 1H),8.43-8.37 (m, 1H), 8.12-8.03 (m, 1H), 7.75-7.66 (m, 1H), 7.49-7.28 (m,4H), 7.28-7.14 (m, 2H), 5.25-5.15 (m, 1H), 4.82-4.67 (m, 2H), 4.28-4.05(m, 4H), 3.06-2.84 (m, 4H). Mixture of rotamers. MS (m/z): 532.2 (M +H)⁺. 23

¹H NMR (400 MHz, DMSO): δ 10.26-9.80 (m, 1H), 8.48-8.31 (m, 1H),7.76-7.68 (m, 1H), 7.64-7.49 (m, 1H), 7.44-7.34 (m, 1H), 7.30-7.02 (m,6H), 6.02-5.86 (m, 1H), 5.59-4.97 (m, 3H), 4.94-4.59 (m, 1H), 4.33-3.95(m, 3H), 1.54-1.30 (m, 3H). Mixture of rotamers. MS (m/z): 565.2 (M +H)⁺. 24

¹H NMR (400 MHz, DMS0): δ 10.42- 10.12 (m, 1H), 8.37-8.08 (m, 1H), 7.90-7.67 (m, 2H), 7.67-7.55 (m, 1H), 7.55- 7.30 (m, 5H), 7.30-7.13 (m, 2H),5.32- 5.18 (m, 1H), 4.90-4.62 (m, 2H), 4.27- 3.91 (m, 4H), 3.11-2.93 (m,2H), 2.93- 2.78 (m, 2H). Mixture of rotamers. 25

¹H NMR (400 MHz, DMSO): δ 10.29 (m, 1H), 8.22 (m, 1H), 7.84 (m, 2H),7.61 (m, 1H), 7.41 (m, 3H), 7.29-7.11 (m, 3H), 5.63-5.20 (m, 3H), 4.79(m, 2H), 4.15 (m, 4H). Mixture of rotamers. 26

¹H NMR (400 MHz, DMSO): δ 10.15-9.81 (m, 1H), 8.77-8.50 (m, 1H),8.07-6.95 (m, 10H), 5.34-5.00 (m, 1H), 4.90-4.47 (m, 2H), 4.12 (s, 4H),2.89-2.73 (m, 2H), 1.21 (s, 6H). Mixture of rotamers. MS (m/z): 559.2(M + H)⁺; r.t. = 1.781 27

MS (m/z): 533.2 (M + H)⁺; r.t. = 1.678 28

NMR (400 MHz, DMSO): δ 12.23 (br s, 1H), 10.01-9.98 (m, 1H), 8.43-8.41(m, 1H), 7.80-7.10 (m, 8H), 7.02 (app d, J = 8.4 Hz, 1H), 5.39-5.30 (m,3H), 4.86-4.70 (m, 3H), 4.13 (app br s, 3H). Mixture of rotamers. MS(m/z): 567.1/569.1 (M + H)⁺ (chlorine isotope pattern).

Example 23-(2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid (Compound 29)

Step 1

To a suspension of the8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-4) (0.20 mmol) in CH₂Cl₂ (1 mL) was added a catalytic amount of DMF(2.5 μL) and dropwise oxalyl chloride (0.60 mmol) at 0° C. The resultingsuspension was warmed up to room temperature and stirred for 1 hr. Thesolvent was removed under vacuum to dryness completely (need to removeoxalyl chloride completely). The obtained residue was dissolved withCH₂Cl₂ (1 mL) and then dropwise into a solution of methyl3-(2-(benzylamino)acetamido)-4-fluorobenzoate (I-19) (0.2 mmol) inCH₂Cl₂ (1 mL) in the presence of DIEA (0.38 mmol). The reaction mixturewas stirred for 15 mins at room temperature and then was charged 50 mLwater. Organic layer was separated and aqueous layer was extracted withCH₂Cl₂ (50 mL). Combined organic layers were washed with H₂O and brinesuccessively, dried over Na₂SO₄, filtered and concentrated to give thecrude product which was purified by column chromatography (0-60% EtOAcin hexanes) to give methyl3-(2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoate(29-Int). ¹H NMR (400 MHz, CDCl₃): δ 9.94 (s, 1H), 9.10-8.85 (m, 1H),7.82 (s, 1H), 7.54-7.30 (m, 6H), 7.26-7.21 (m, 1H), 7.21-7.10 (m, 1H),7.01 (d, J=8.3 Hz, 1H), 5.64-4.45 (m, 5H), 4.36-4.06 (m, 4H), 3.95 (s,3H). Mixture of rotamers. MS (m/z): 563.1 (M+H)⁺.

Step 2

Methyl3-(2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoate(29-Int) (0.14 mmol) was dissolved in THF/MeOH/H₂O (3:2:1, 5.0 mL) andfollowed by addition of LiOH monohydrate (0.85 mmol). The reactionmixture was stirred at room temperature for 2 hrs. Solvents were removedunder vacuum and acidified to pH=3.0 by addition of 3N HCl. Solid wascollected and dried to give3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-5-fluorobenzoicacid. ¹H NMR (400 MHz, DMSO): δ 10.11-9.96 (m, 1H), 8.67-8.53 (m, 1H),7.78-7.66 (m, 2H), 7.43-7.26 (m, 7H), 7.11-7.03 (m, 1H), 5.48-4.10 (m,9H). Mixture of rotamers. MS (m/z): 549.1 (M+H)⁺.

The following compounds were prepared according to the proceduresdescribed in Example 2 using the appropriate intermediates.

Compound Structure Characterization Data 30

¹H NMR (400 MHz, DMSO): δ 10.17-10.01 (m, 1H), 8.64-8.49 (m, 1H),7.75-7.67 (m, 2H), 7.48-7.30 (m, 4H), 7.26-7.17 (m, 2H), 7.09-7.02 (m,1H), 5.43-4.12 (m, 9H). Mixture of rotamers. MS (m/z): 563.2 (M + H)⁺.31

¹H NMR (400 MHz, DMSO): δ 10.26-10.18 (m, 1H), 8.27-8.19 (m, 1H),7.82-7.76 (m, 1H), 7.74-7.66 (m, 1H), 7.65-7.59 (m, 1H), 7.47-7.40 (m,1H), 7.40-7.26 (m, 6H), 7.10- 7.00 (m, 1H), 5.49-4.04 (m, 9H). Mixtureof rotamers. MS (m/z): 531.1 (M + H)⁺. 32

¹H NMR (400 MHz, DMSO): δ 10.10-9.98 (m, 1H), 8.67-8.53 (m, 1H),7.76-7.67 (m, 1H), 7.61-7.51 (m, 1H), 7.42-7.34 (m, 5H), 7.34- 7.26 (m,1H), 7.19-7.11 (m, 1H), 7.10- 7.03 (m, 1H), 5.47-4.05 (m, 9H). Mixtureof rotamers. MS (m/z): 533.1 (M + H)⁺. 33

¹H NMR (400 MHz, DMSO): δ 10.11-9.97 (m, 1H), 8.68-8.52 (m, 1H),7.78-7.66 (m, 2H), 7.42-7.34 (m, 5H), 7.34-7.25 (m, 2H), 7.14- 6.99 (m,2H), 5.46-4.09 (m, 9H). Mixture of rotamers. MS (m/z): 515.1 (M + H)⁺.

Example 2A3-(2-(9-chloro-N-(2-fluorobenzyl)-1-methyl-4,5-dihydro-1H-benzo[2,3]oxepino[4,5-]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid (Compound 34)

Step 1

To a suspension of the9-chloro-1-methyl-4,5-dihydro-1H-benzo[2,3]oxepino[4,5-c]pyrazole-3-carboxylicacid (1.8 mmol) in CH₂Cl₂ (10 mL) was added catalytic amount of DMF (25uL) and dropwise oxalyl chloride (8.9 mmol) at 0° C. The resultingsuspension was warmed up to room temperature and stirred for 1 hr. Thesolvent was removed under vacuum to dryness completely. The obtainedresidue was dissolved with CH₂Cl₂ (10 mL) and then dropwise into asolution of methyl4-fluoro-3-(2-((2-fluorobenzyl)amino)acetamido)benzoate (I-17) (1.8mmol) in CH₂Cl₂ (10 mL) in the presence of DIEA (3.6 mmol). The reactionmixture was stirred for 30 mins at room temperature and then was chargedwith 50 mL water. Organic layer was separated and aqueous layer wasextracted with CH₂Cl₂ (50 mL). Combined organic layers were washed withH₂O and brine successively, dried over Na₂SO₄, filtered and concentratedto give the crude product which was purified by column chromatography(0-60% EtOAc in hexanes) to give methyl3-(2-(9-chloro-N-(2-fluorobenzyl)-1-methyl-4,5-dihydro-1H-benzo[2,3]oxepino[4,5-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoate.¹H NMR (400 MHz, CDCl₃): δ 10.11 (s, 1H), 8.74 (s, 1H), 7.72 (s, 1H),7.38-7.31 (m, 3H), 7.27-7.22 (m, 1H), 7.12-6.97 (m, 3H), 5.28-4.79 (m,2H), 4.38 (t, J=6.4 Hz, 3H), 4.05-3.94 (m, 4H), 3.84 (s, 3H), 3.13 (t,J=6.4 Hz, 2H). MS (m/z): 595.1 (M+H)⁺.

Step 2

Methyl3-(2-(9-chloro-N-(2-fluorobenzyl)-1-methyl-4,5-dihydro-1H-benzo[2,3]oxepino[4,5-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoate(1.6 mmol) was dissolved in THF/MeOH/H₂O (3:2:1, 10 mL) and followed byaddition of LiOH monohydrate (9.7 mmol). The reaction mixture wasstirred at room temperature for 4 hr and diluted with 10 ml of water andacidified to pH=3.0. Solid was collected and dried to yield3-(2-(9-chloro-N-(2-fluorobenzyl)-1-methyl-4,5-dihydro-1H-benzo[2,3]oxepino[4,5-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid. ¹H NMR (400 MHz, DMSO): δ 10.03 (s, 1H), 8.69-8.48 (m, 1H),7.84-7.66 (m, 2H), 7.52-7.30 (m, 4H), 7.28-7.12 (m, 3H), 5.18-4.20 (m,6H), 4.05-3.92 (m, 3H), 3.11-2.93 (m, 2H). Mixture of rotamers. MS(m/z): 581.1 (M+H)⁺.

Example 33-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid (Compound 35)

To a 40 mL reaction vessel was added reagents in the following order:2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)aceticacid (I-22) (0.10 mmol), THF (2 mL), and N-methyl morpholine (1.4 mmol).This was stirred until complete dissolution. Next was added2-chloro-4,6-dimethoxy-1,3,5-triazine (0.20 mmol) and this solution wasstirred for 20 mins at 50° C. until a white precipitate fully formed.The precipitate was physically agitated to ensure all solids were wellstirred. Next was added methyl 3-aminobenzoate (0.30 mmol) and thereaction was stirred for 30 mins at 50° C. and then diluted with 5 mL ofMeOH and then directly purified by acetic acid modified (0.05%)reverse-phase chromatography (30 to 80%, ACN/water). Ester intermediatewas used without further purification and was dissolved in 2 mL of MeOH,2 mL of THF, and treated with 1 mL of 1 N KOH aqueous solution (1.0mmol). After heating the homogenous solution to 60° C. for 30 mins thereaction was cooled to room temperature, quenched with AcOH (17 N,approx 0.06 mL) to about pH-6. The reaction was diluted with ethylacetate (30 mL), rinsed with 2×5 mL of water, and the ethyl acetatefraction was partially concentrated to give3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid as a white solid. ¹H NMR (400 MHz, DMSO): δ 11.91 (br s, 1H), 10.15(br s, 1H), 8.26-8.20 (m, 1H), 7.85-7.62 (m, 3H), 7.48-7.29 (m, 4H),7.20 (app q, J=7.5 Hz, 2H), 7.05 (dd, J=1.7, 8.7 Hz, 1H), 5.39-5.35 (m,2H), 4.84-4.70 (m, 2H), 4.23-4.10 (m, 5H). Mixture of rotamers. MS(m/z): 549.2/551.2 (M+H)⁺ (chlorine isotope pattern).

Example 43-(2-(8-chloro-N-(cyclopentylmethyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid (Compound 36)

To a 40 mL reaction vessel was added reagents in the following order:8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-4) (0.10 mmol, THF (3 mL) and N-methyl morpholine (1.0 mmol). Thiswas stirred until complete dissolution. Next was added2-chloro-4,6-dimethoxy-1,3,5-triazine (0.11 mmol) and this solution wasstirred for 20 mins at 50 C until a white precipitate fully formed. Theprecipitate was physically agitated with rapid stirring to ensure allsolids were well mixed. Next was added methyl3-(2-((cyclopentylmethyl)amino) acetamido)-4-fluorobenzoate (I-70) (0.10mmol) and the reaction was stirred for 30 mins at 50° C. and thendiluted with 5 mL of MeOH and then purified by acetic acid modified(0.05%) reverse phase chromatography (30 to 80%, water/ACN). Theintermediate ester was carried without further manipulation and wasdissolved in 2 mL of MeOH, 2 mL of THF, and treated with 1 mL of 1 N KOHaqueous solution (1.0 mmol). After heating the homogenous solution to60° C. for 30 mins the reaction was cooled to room temperature, quenchedwith AcOH (17 N, approx. 0.06 mL) to about pH=6. The reaction wasdiluted with 30 mL of ethyl acetate, rinsed with 2×5 mL of water, andthe ethyl acetate fraction was partially concentrated to give3-(2-(8-chloro-N-(cyclopentylmethyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid as a white solid. ¹H NMR (400 MHz, DMSO): δ 10.06-10.01 (m, 1H),8.46-8.40 (m, 1H), 7.82-7.78 (m, 2H), 7.36-7.32 (m, 2H), 7.03-7.00 (m,1H), 5.32-5.20 (m, 2H), 4.89 (br s, 1H), 4.29 (br s, 2H), 4.24-3.90 (m,4H), 2.10-2.05 (m, 1H), 1.87-1.67 (m, 6H), 1.09-1.02 (m, 2H). Mixture ofrotamers. MS (m/z): 541.2/542.2 (M+H)⁺ (chlorine isotope pattern).

Example 53-(2-(8-chloro-N-(cyclopentylmethyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid (Compound 37)

3-(2-(8-chloro-N-(cyclopentylmethyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid was made following the procedures described in Example 4, exceptfor the substitution of methyl3-(2-((cyclopentylmethyl)amino)acetamido)benzoate (I-71) in the place ofmethyl 3-(2-((cyclopentylmethyl)amino)acetamido)-4-fluorobenzoate(I-70). ¹H NMR (400 MHz, DMSO): δ 12.66 (br s, 1H), 10.18-10.14 (m, 1H),8.23 (app d, J=10.0 Hz, 1H), 7.89-7.12 (m, 5H), 7.06-7.02 (m, 1H),5.35-5.25 (m, 2H), 4.89 (br s, 1H), 4.72 (br s, 2H), 4.43 (br s, 1H),4.14-4.00 (m, 3H), 1.99-0.96 (m, 9H). Mixture of rotamers. MS (m/z):523.2/525.2 (M+H)⁺ (chlorine isotope pattern).

Example 63-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-5-fluorobenzoicacid (Compound 38)

Step 1

To a mixture of2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)aceticacid (I-22) (0.032 mmol), DIEA (0.16 mmol) and ethyl3-amino-5-fluorobenzoate (0.05 mmol) in EtOAc (0.5 mL) was added T₃P(50% wt in EtOAc, 0.5 mL) and stirred at room temperature for 30 mins.Purification by HPLC yielded ethyl3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-5-fluorobenzoate(38-Int). ¹H NMR (400 MHz, CDCl₃): δ 9.38-8.71 (m, 1H), 7.95-7.69 (m,1H), 7.59 (s, 1H), 7.49-7.39 (m, 3H), 7.26-7.22 (m, 1H), 7.14 (td,J=1.1, 7.5 Hz, 1H), 7.08-7.02 (m, 1H), 7.00 (d, J=8.7 Hz, 1H), 5.53 (s,2H), 5.08-4.81 (m, 1H), 4.67-4.47 (m, 1H), 4.40 (q, J=7.1 Hz, 2H), 4.17(s, 3H), 1.42 (t, J=7.1 Hz, 3H). MS (m/z): 595.1 (M+H)⁺.

Step 2

Ethyl3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-5-fluorobenzoate(38-Int) (8 mg, 0.0134 mmol) was dissolved in THF/MeOH/H₂O (3:2:1, 5.0mL) and followed by addition of LiOH monohydrate (0.08 mmol). Thereaction mixture was stirred at room temperature for 2 hrs. The solventswere removed under vacuum and acidified to pH=3.0 by addition of 3N HClto give3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-5-fluorobenzoicacid as a solid. ¹H NMR (400 MHz, DMSO): δ 10.59-10.47 (m, 1H), 7.99 (s,1H), 7.87-7.79 (m, 1H), 7.79-7.72 (m, 1H), 7.53-7.43 (m, 1H), 7.43-7.36(m, 3H), 7.32-7.23 (m, 2H), 7.14-7.08 (m, 1H), 5.51-4.13 (m, 9H).Mixture of rotamers. MS (m/z): 567.0 (M+H)⁺.

The following compounds were prepared according to the proceduresdescribed in Example 6 using the appropriate intermediates.

Compound Structure Characterization Data 39

MS (m/z): 563.1 (M + H)⁺; r.t. = 1.886

Example 73-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-2-methylpropanoicacid (Compound 40)

Step 1

To a mixture of2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)aceticacid (I-22) (0.15 mmol), DIEA (0.45 mmol) and HATU (0.18 mmol) in DMF (1mL) was added ethyl 3-amino-2-methylpropanoate (0.22 mmol) and stirredat room temperature for 10 mins. Then the reaction was diluted with 20mL water and extracted with EtOAc (20 mL, twice). Combined organiclayers were washed with H₂O and brine successively, dried over Na₂SO₄,filtered and concentrated to give the crude product which was purifiedby column chromatography (0-80% EtOAc in hexanes) to yield ethyl3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-2-methylpropanoate(40-Int). ¹H NMR (400 MHz, CDCl₃): δ 7.47 (d, J=2.4 Hz, 1H), 7.43-7.33(m, 1H), 7.33-7.29 (m, 1H), 7.22 (dd, J=2.4, 8.7 Hz, 1H), 7.14 (dt,J=3.8, 7.5 Hz, 1H), 7.08 (t, J=9.2 Hz, 1H), 6.98 (d, J=8.7 Hz, 1H),5.60-4.74 (m, 5H), 4.25-3.89 (m, 6H), 3.62-3.39 (m, 1H), 3.40-3.24 (m,1H), 2.72-2.61 (m, 1H), 1.23-1.14 (m, 6H). Mixture of rotamers. MS(m/z): 543.1 (M+H)⁺.

Step 2

Ethyl3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-2-methylpropanoate(40-Int) (0.125 mmol) was dissolved in THF/MeOH/H₂O (3:2:1, 5.0 mL) andfollowed by addition of LiOH monohydrate (0.75 mmol). The reactionmixture was stirred at room temperature for 2 hr. The solvents wereremoved under vacuum then water (5 mL) was added and acidified to pH=3.0by addition of 3N HCl to give3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-2-methylpropanoicacid. ¹H NMR (400 MHz, DMSO): δ 12.26 (s, 1H), 8.10-7.99 (m, 1H),7.73-7.67 (m, 1H), 7.40-7.29 (m, 3H), 7.24-7.16 (m, 2H), 7.09-7.02 (m,1H), 5.43-3.89 (m, 9H), 3.32-3.06 (m, 2H), 1.06-0.97 (m, 3H). Mixture ofrotamers. MS (m/z): 515.0 (M+H)⁺.

The following compounds were prepared according to the proceduresdescribed in Example 7 using the appropriate intermediates.

Compound Structure Characterization Data 41

¹H NMR (400 MHz, CDCl₃): δ 7.35 (s, 1H), 7.33-7.24 (m, 1H), 7.15-7.08(m, 1H), 7.08- 6.94 (m, 2H), 6.94-6.80 (m, 2H), 5.45- 3.93 (m, 9H),3.33-3.19 (m, 2H), 1.15-1.05 (m, 6H). Mixture of rotamers. MS (m/z):529.1 (M + H)⁺. 42

¹H NMR (400 MHz, DMSO): δ 7.99-7.88 (m, 1H), 7.73-7.68 (m, 1H),7.39-7.29 (m, 3H), 7.23-7.14 (m, 2H), 7.08-7.03 (m, 1H), 5.42- 5.26 (m,3H), 4.66-4.55 (m, 2H), 4.19- 3.89 (m, 4H), 3.32-3.27 (m, 2H), 1.03-0.73(m, 4H). Mixture of rotamers. MS (m/z): 527.3 (M + H)⁺.

Example 84-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid (Compound 43)

Step 1

To8-chloro-N-(2-fluorobenzyl)-N-(2-hydroxyethyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamide(I-73) (0.45 mmol) was added THF (7 mL) and methyl3-fluoro-4-hydroxybenzoate (0.88 mmol), triphenyl phosphine solid (0.90mmol) the resulting solution cooled to 0° C. and lastly DIAD(diisopropyl azodicarboxylate) dropwise (0.20 mL, Aldrich™ commercialstock at 95% weight by weight content, 0.96 mmol). The internaltemperature of the reaction maintained at 0° C. for 10 mins, thenallowed to warm to room temperature over 20 mins. After a full hr atroom temperature, the reaction was diluted with THF (5 mL), filtered,and directly purified by TFA modified (0.05%) reverse phasechromatography (40 to 90%, water/ACN). All fractions were reduced todryness under vacuum and subjected to a free base event using polymerimmobilized carbonate (SPE-CO3H Varian cartridge, 0.90 nominal load withMeOH mobilizer, 10 mL) to give the methyl ethyl intermediate as an offwhite powder (43-Int). ¹H NMR (400 MHz, DMSO): δ 7.78-7.69 (m, 2H), 7.63(br s, 1H), 7.42-7.38 (m, 3H), 7.18 (app dt, J=12.3, 8.0 Hz, 3H), 7.04(app d, J=8.0 Hz, 1H), 5.40-5.35 (m, 2H), 5.29 (br s, 1H), 4.83 (br s,1H), 4.52-4.38 (m, 2H), 4.15-4.10 (m, 2H), 3.93 (s, 3H) 3.73 (s, 3H).Mixture of rotamers. MS (m/z): 568.2/570.2 (M+H)⁺ (chlorine isotopepattern).

Step 2

The ester, methyl4-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoate(43-Int), generated in the previous step was then dissolved in THF (10mL), MeOH (2 mL), and 1.0 M KOH (2.0 mmol). The resulting homogenoussolution was heated to 60° C. for 2 hrs. At this time the reaction wascooled back to room temperature, quenched with 0.12 mL of AcOH (2 mmol,to PH-6 using Whatman-4 color-strip indicator paper to monitor). Thereaction was then diluted with water (20 mL) and extracted with ethylacetate (3×100 mL). The organic extracts were further washed with water(2×15 mL). The organic extracts were removed to dryness and allowed toprecipitate from MeOH/water (10 mL, 9:1) to give4-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid as a white solid. ¹H NMR (400 MHz, DMSO): δ 12.51 (br s, 1H),7.89-7.19 (m, 9H), 7.07 (dd, J=1.7, 8.7 Hz, 1H), 5.30-5.15 (m, 3H), 4.85(br s, 1H), 4.43-4.18 (m, 3H, 4.04-3.99 (m, 3H), 3.78 (m, 1H). Mixtureof rotamers. MS (m/z): 554.1/556.1 (M+H)⁺ (chlorine isotope pattern).

The following compounds were prepared according to the proceduresdescribed in Example 8 using the appropriate intermediates.

Compound Structure Characterization Data 44

¹H NMR (400 MHz, DMSO): δ 12.56 (br s, 1H), 7.79-7.10 (m, 10 H), 7.06(dd, J = 1.7, 8.7 Hz, 1H), 5.40-5.25 (m, 3H), 4.82 (br s, 1H), 4.33-4.13(m, 3H), 4.04 (br s, 3H), 3.75 (m, 1H). Mixture of rotamers. MS (m/z):536.1/538.1 (M + H)⁺ (chlorine isotope pattern). 45

MS (m/z): 536.2/538.2 (M + H)⁺ (chlorine isotope pattern); r.t. = 1.60846

MS (m/z): 550.2/552.2 (M + H)⁺ (chlorine isotope pattern); r.t. = 1.63247

MS (m/z): 564.2/566.2 (M + H)⁺ (chlorine isotope pattern); r.t. = 1.75948

MS (m/z): 554.2/556.2 (M + H)⁺ (chlorine isotope pattern); r.t. = 1.75049

¹H NMR (400 MHz, DMSO): δ 12.50 (br s, 1H), 7.82 (app d, J = 8.0 Hz,1H), 7.75 (br s, 2H), 7.30 (app d, J = 8.0 Hz, 1H), 7.03- 7.00 (m, 1H),5.40-5.30 (m, 2H), 4.30 (br s, 1H), 4.22-3.88 (m, 6H), 3.82-3.78 (m,1H), 3.56-3.54 (m, 1H), 2.13-2.10 (m, 1H), 2.06 (s, 3H), 2.02 (s, 3H),1.69-1.38 (m, 6 H), 1.24-1.15 (m, 2H). Mixture of rotamers. MS (m/z):538.1/540.1 (M + H)⁺ (chlorine isotope pattern). 50

MS (m/z): 572.2/574.2 (M + H)⁺ (chlorine isotope pattern); r.t. = 1.69051

MS (m/z): 604.3/606.3 (M + H)⁺ (chlorine isotope pattern); r.t. = 1.76052

¹H NMR (400 MHz, d₄-MeOH): δ 7.59- 7.15 (m, 9H), 6.78-6.73 (m, 1H),5.38- 5.24 (m, 3H), 4.85 (br s, 1H, partly obscured), 4.42-4.21 (m, 3H),4.16-4.04 (m, 3H), 3.69 (app t J = 8.2 Hz, 1H). Mixture of rotamers. MS(m/z): 554.1/ 556.1 (M + H)⁺ (chlorine isotope pattern). 53

MS (m/z): 556.2 (M + H)⁺; r.t. = 1.650 54

¹H NMR (400 MHz, d₄-MeOH): δ 7.52- 6.83 (m, 10H), 5.38-5.20 (m, 3H),4.83 (br s, 1H), 4.43-3.87 (m, 7H). Mixture of rotamers. MS (m/z): 538.1(M + H)⁺.

Example 94-(2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid (Compound 55)

Step 1

To a solution of8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-4) (0.16 mmol) in anhydrous DMF (1 mL) were added HATU (0.18 mmol),DIEA (0.48 mmol) and methyl 4-(2-(benzylamino)ethoxy)-3-fluorobenzoate(I-52) (0.16 mmol) and the resulting mixture was stirred at roomtemperature until completion (2 hrs). The reaction mixture was thendiluted with water (10 mL) and extracted with ethyl acetate (20 mL×3).The organic phase was washed with 1N HCl, water, brine, dried overNa₂SO₄ and concentrated in vacuum. The crude residue was triturated withMeOH to afford methyl4-(2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoateas white solid (55-Int). MS (m/z): 550.1/552.1 (M+H)⁺ (chlorine isotopepattern).

Step 2

A suspension of4-(2-(N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid (61-int) (20 mg, 0.036 mmol) in a 3:2:1 THF-MeOH-H₂O solution (2mL) was treated with a 6M solution of LiOH (100 uL) and the resultingmixture was stirred at 35° C. until complete conversion (3 hrs). Thereaction mixture was then concentrated under reduced pressure. The cruderesidue was dissolved in water and 50% acetic acid was added until pH 5at which point a white solid separated out. The solid was collected byfiltration, washed with water and dried under high vacuum to give4-(2-(N-benzyl-8-chloro-1-methyl-1,4dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)-3-fluorobenzoicacid. ¹H NMR (400 MHz, DMSO): δ 7.75-6.98 (m, 11H), 5.44-5.30 (m, 5H),4.45-3.99 (m, 6H). Mixture of rotamers. MS (m/z): 536.2/538.2 (M+H)⁺(chlorine isotope pattern).

The following compounds were prepared according to the proceduresdescribed in Example 9 using the appropriate intermediates.

Compound Structure Characterization Data 56

MS (m/z): 520.1 (M + H)⁺; r.t. = 1.636 57

MS (m/z): 538.1 (M + H)⁺; r.t. = 1.628 58

¹H NMR (400 MHz, DMSO): δ 7.75-7.03 (m, 11H), 5.41-5.26 (m, 2H), 4.81(s, 1 H), 4.44-4.34 (m, 3H), 4.20-3.98 (m, 4H), 3.82-3.65 (m, 1H).Mixture of rotamers. MS (m/z): 538.1 (M + H)⁺ 59

¹H NMR (400 MHz, DMSO): δ 7.85-7.07 (m, 10H), 5.43-5.26 (m, 2H), 4.81(s, 1H), 4.44-4.28 (m, 3H), 4.14-3.96 (m, 4H), 3.79-3.68 (m, 1H).Mixture of rotamers. MS (m/z): 556.1 (M + H)⁺ 60

MS (m/z): 554.1/556.1 (M + H)⁺ (chlorine isotope pattern); r.t. = 1.667

Example 103-fluoro-4-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)benzoicacid (Compound 61)

Step 1

HATU (0.22 mmol) was added to a mixture of8-fluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-5) (0.20 mmol), N,N-diisopropylamine (0.44 mmol) and DMF (2 mL) atroom temperature. After stirring for 20 mins, amine (I-56) (0.20 mmol)in DMF (1 mL) was added and the reaction mixture was stirred for 4 hrsat room temperature. The mixture was diluted with H₂O (10 mL) and EtOAc(10 mL), the layers were separated, and the H₂O layer was washed withEtOAc (×2, 10 mL). The combined organic extracts were washed with H₂O(10 mL), brine (10 mL), and then dried (MgSO₄). After removal ofsolvent, the crude material was purified by chromatography (solid load,silica gel, 0-60% EtOAc/hexanes) to give methyl3-fluoro-4-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)benzoate(61-Int) as a white solid. ¹H NMR (400 MHz, CDCl₃): δ 7.84-7.64 (m, 2H),7.43-7.29 (m, 1H), 7.21-7.13 (m, 1H), 7.13-7.02 (m, 2H), 7.01-6.87 (m,3H), 5.60-5.54 (m, 1H), 5.50-5.43 (m, 2H), 5.30 (s, 3H), 5.01-4.94 (m,1H), 4.45-4.37 (m, 2H), 4.37-4.30 (m, 1H), 4.09-4.06 (m, 3H), 3.91-3.84(m, 2H).

Step 2

KOH (1.0 M in H₂O, 1.0 mmol) was added to a solution of methyl3-fluoro-4-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)benzoate (61-Int) (0.2 mmol) in THF (4.0 mL) and MeOH (1.0 mL) and thesolution was stirred at 50° C. for 2 hrs. After removal of solvent(aspirator), the crude residue was diluted with water (10 mL) and thesolution was acidified with acetic acid (to pH ˜5). The resulting whiteprecipitate was collected by vacuum filtration, washed with H₂O (20 mL),and dried overnight on high vacuum to give3-fluoro-4-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethoxy)benzoicacid. ¹H NMR (400 MHz, DMSO): δ 7.75-7.64 (m, 1H), 7.57-7.01 (m, 9H),5.51-5.23 (m, 3H), 4.89-4.80 (m, 1H), 4.49-4.29 (m, 3H), 4.13-3.98 (m,3H), 3.83-3.72 (m, 1H). MS (m/z): 538.2 (M+H)⁺. Mixture of rotamers.

The following compound was prepared according to the proceduresdescribed in Example 10 using the appropriate intermediates.

Compound Structure Characterization Data 62

¹H NMR (400 MHz, DMSO): δ 7.82-7.12 (m, 9H), 5.51-5.24 (m, 3H),4.87-4.78 (m, 1H), 4.48-4.30 (m, 3H), 4.11-3.96 (m, 3H), 3.81- 3.72 (m,1H). Mixture of rotamers. MS (m/z): 556.2 (M + H)⁺.

Example 113-((2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid (Compound 63)

Step 1

To a 40 mL reaction vessel was charged8-chloro-N-(2-fluorobenzyl)-N-(2-hydroxyethyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamide(I-73) (0.10 mmol) and CH₂Cl₂ (10 mL) followed by the addition ofcommercial Dess-Martin Periodinane (Sigma-Aldrich,1,1,1-Tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)-one, 0.20 mmol)and buffered with solid sodium bicarbonate (0.20 mmol). The milky whitesuspension was rapidly stirred for 2 hrs and then the reaction was thendirectly diluted with ethyl acetate (150 mL) and washed with water (3×15mL). The resulting organic extract was concentrated to a residue (invacuo) and used directly without delay or further manipulation. Theresidue was dissolved in 9:1 MeOH/AcOH (5 mL) and treated withmethyl-3-amino benzoate (0.40 mmol). The resulting reaction was thenstirred at room temperature for 30 mins. and then treated with sodiumcyanoborohydride (1.0 mmol, portion-wise over 30 mins.) followed bystirring at room temperature for an additional 1 hr. The resultingreaction was diluted with ethyl acetate (150 mL) and washed with water(3×25 mL). The resulting organic extracts were concentrated in vacuo andthen directly subjected to reverse phase chromatography usingTFA-modified (0.05%) water/ACN (35 to 80%). All fractions were reducedto dryness under vacuum and subjected to a free base event using polymerimmobilized carbonate (SPE-COSH Varian cartridge, 0.90 nominal load withMeOH mobilizer, 10 mL) to give methyl3-((2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoate (63-Int) as a white solid. ¹H NMR (400 MHz, d₄-MeOH): δ7.48-7.19 (m, 8H), 7.03-6.92 (3H), 7.18 (app d, J=8.0 Hz, 1H), 6.62 (appdt, J=12.2, 8.0 Hz, 1H), 5.30-5.20 (m, 2H), 4.83 (br s, 2H), 4.21 (t,J=5.0 Hz, 2H), 3.85 (s, 3H), 3.74 (s, 3H), 3.38 (t, J=5.0 Hz, 2H).Mixture of rotamers. MS (m/z): 549.2/551.2 (M+H)⁺ (chlorine isotopepattern).

Step 2

Methyl3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoate (63-Int) (0.075 mmol) was dissolved in THF (3 mL),MeOH (2 mL), and 1.0 M KOH (1.0 mmol). The resulting homogenous solutionwas heated to 60° C. for 2 hrs. At this time the reaction was cooledback to room temperature, quenched with AcOH (1.2 mmol, to pH=6 usingWhatman-4 color-strip indicator paper to monitor). The reaction was thendiluted with water (5 mL) and extracted with ethyl acetate (3×20 mL).The organic extracts were further washed with water (2×5 mL). Theorganic extracts were removed to dryness and allowed to precipitate fromMeOH/water (3 mL, 9:1) to give3-((2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoic acid as a white solid. ¹H NMR (400 MHz, d₄-MeOH): δ 7.48-7.10(m, 7H), 7.02-6.90 (m, 3H), 6.61 (br d, J=8.0 Hz, 1H), 5.40-5.20 (m,2H), 4.80 (br s, 2H), 4.21 (t, J=7.5 Hz, 2H), 3.91 (br s, 3H), 3.38 (t,J=7.5 Hz, 2H). Mixture of rotamers. MS (m/z): 535.2/537.2 (M+H)⁺(chlorine isotope pattern).

Example 123-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid (Compound 64)

3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)-4-fluorobenzoicacid was prepared according to the procedures described in Example 3 for3-(2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid, using methyl 3-amino-4-fluorobenzoate in place of methyl-3-aminobenzoate in step 1. MS (m/z): 567.2/569.1 (M+H)⁺ (chlorine isotopepattern); r.t.=1.734.

Example 133-((2-(8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid (Compound 65)

Step 1

HATU (1.2 mmol) was added to a mixture of8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-4) (1.1 mmol), Hunigs Base (2.4 mmol) and DMF (5 mL). A solution ofmethyl 4-fluoro-3-((2-((3-fluorobenzyl)amino)ethyl)amino) benzoate(I-46) (1.1 mmol) in DMF (3 mL) was added and the reaction mixturestirred at room temperature for 4 hrs. The mixture was diluted withwater and ethyl acetate. The layers were separated and the aqueous phasewas washed with ethyl acetate. The combined organic extracts were washedwith water, brine, and then dried over MgSO₄. The material was purifiedby chromatography (silica, 0-60% ethyl acetate/hexanes) to give methyl3-((2-(8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoate(65-Int). ¹H NMR (400 MHz, DMSO): δ 7.61-7.29 (m, 2H), 7.23-6.86 (m,8H), 6.20-5.89 (m, 1H), 5.45-4.65 (m, 4H), 4.14-3.95 (m, 4H), 3.84-3.77(m, 3H), 3.58-3.46 (m, 1H), 3.46-3.37 (m, 2H).

Step 2

1N KOH (4 mmol, 5 equiv.) was added to a solution of methyl3-((2-(8-chloro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoate(65-Int) (0.80 mmol) in THF (4 mL) and MeOH (2 mL) and the solution wasstirred at 50° C. for 2 hrs. The solvent was removed and the cruderesidue was diluted with water. The aqueous solution was acidified withacetic acid (pH ˜5) to give the title compound as a white precipitate.¹H NMR (400 MHz, DMSO): δ 7.61-6.76 (m, 10H), 6.21-5.77 (m, 1H),5.46-3.94 (m, 9H), 3.56-3.38 (m, 2H). Mixture of rotamers. MS (m/z):537.2 (M+H)⁺.

The following compounds were prepared according to the proceduresdescribed in Example 13 using the appropriate intermediates.

Compound Structure Characterization Data 66

¹H NMR (400 MHz, DMSO): δ 7.77-7.52 (m, 1H), 7.38-6.81 (m, 9H),6.12-5.82 (m, 1H), 5.49-3.94 (m, 9H), 3.40 (s, 2H), 2.25 (s, 3H).Mixture of rotamers. MS (m/z): 549.3/551.3 (M + H)⁺ (chlorine isotopepattern). 67

¹H NMR (400 MHz, DMSO): δ 7.12 (m, 11H), 6.14-5.81 (m, 1H), 5.47-3.82(m, 9H), 3.40 (s, 2H), 2.25 (s, 3H). Mixture of rotamers. MS (m/z):533.3 (M + H)⁺. 68

¹H NMR (400 MHz, DMSO): δ 7.88-7.60 (m, 1H), 7.37-6.84 (m, 8H),6.20-5.90 (m, 1H), 5.48-3.76 (m, 9H), 3.52-3.37 (m, 2H), 2.25 (s, 3H).Mixture of rotamers. MS (m/z): 551.3 (M + H)⁺. 69

¹H NMR (400 MHz, DMSO): δ 7.56-6.75 (m, 10H), 5.93 (m, 1H), 5.43-5.09(m, 2H), 4.76 (s, 1H), 4.20-3.90 (m, 5H), 3.56 (m, 1H), 3.40 (dd, J =16.7 Hz, 2H). Mixture of rotamers. MS (m/z): 537.3 (M + H)⁺. 70

¹H NMR (400 MHz, DMSO): δ 7.77-6.72 (m, 11H), 6.09-5.81 (m, 1H),5.41-5.21 (m, 2H), 4.81-4.71 (m, 1H), 4.26-3.86 (m, 5H), 3.61-3.53 (m,1H), 3.46-3.37 (m, 2H). Mixture of rotamers. MS (m/z): 519.2 (M + H)⁺.71

MS (m/z): 519.2 (M + H)⁺; r.t = 1.812 72

MS (m/z): 535.1/537.1 (M + H)⁺ (chlorine isotope pattern); r.t = 1.77173

¹H NMR (400 MHz, DMSO): δ 12.62 (br s, 1H), 7.92-7.23 (m, 7H), 6.89 (appd, J = 7.4 Hz, 1H), 6.78 (app t, J = 7.8 Hz, 1H), 5.58 (br s 1H, NH),5.30-5.20 (m, 2H), 4.89 (br s, 2H), 4.33-3.97 (m, 5H), 3.62-3.54 (m,2H). Mixture of rotamers. MS (m/z): 555.2 (M + H)⁺. 74

MS (m/z): 537.1 (M + H)⁺; r.t = 1.698

Example 144-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid (Compound 75)

Step 1

HATU (43.1 mmol) (Oakwood) was added to a solution of the8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-4) (39 mmol) and DIEA (15 mL) (Aldrich) in DMF (300 mL) at roomtemperature. After stirring for 20 mins, methyl4-((benzylamino)methyl)benzoate (I-23) (39 mmol) was added neat as anoil followed by a rinse of DMF (20 mL) and the reaction mixture wasstirred for 2 hrs. The mixture was poured onto ice and the resultingprecipitate was filtered, rinsed with H₂O (˜200 mL). The solid wasdissolved in DCM (˜300 mL), washed with NaHCO₃ (sat.), H₂O and dried(MgSO₄). The crude material was purified by chromatography (silica gel,loaded neat with DCM rinse, 10-100% EtOAc/Hexanes) and after removal ofsolvent, an oil was obtained. The oil was suspended in EtOH (˜100 mL)and the mixture was heated to reflux while stirring. After stirring for1 hr, the mixture was cooled to room temperature and the resulting solidwas filtered and rinsed with cold EtOH (˜50 mL). The filtrate wasconcentrated (aspirator) and the crystallization was repeated to givemethyl4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoate.¹H NMR (400 MHz, DMSO): δ 7.99-7.87 (m, 2H), 7.75-7.65 (m, 1H),7.45-7.22 (m, 8H), 7.12-7.02 (m, 1H), 5.48-5.41 (m, 2H), 5.30-5.21 (m,2H), 4.64-4.55 (m, 2H), 4.19-4.05 (m, 3H), 3.85 (s, 3H). MS (m/z):502.0/504.0 (M+H)⁺ (chlorine isotope pattern).

Step 2

1.0 M potassium hydroxide (53 mmol) was added to a solution of methyl4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoate(17.5 mmol) in THF (56 mL) and methanol (14 mL) and the mixture washeated to 50° C. for 2 hr. After cooling to room temperature, thesolvent was removed (aspirator). The crude residue was diluted withwater and the aqueous solution was acidified with acetic acid (to pH ˜6)resulting in a precipitate. The precipitate was collected by vacuumfiltration and dried overnight under high vacuum. The solid wascollected and EtOH (125 mL) was added. The mixture was stirred at 85° C.for 2 hrs and then cooled to room temperature. The solid was filtered,rinsed with cold EtOH (˜75 mL) and dried overnight under high vacuum togive4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid. ¹H NMR (400 MHz, DMSO): δ 7.96-7.85 (m, 2H), 7.74-7.67 (m, 1H),7.42-7.20 (m, 8H), 7.12-6.97 (m, 1H), 5.46-5.42 (m, 2H), 5.28-5.21 (m,2H), 4.62-4.49 (m, 2H), 4.18-4.07 (m, 3H). Mixture of rotamers. MS (m/z)488.2/490.2 (M+H)⁺ (chlorine isotope pattern).

Methanol (400 mL) was added to a mixture of4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid (8.20 mmol) and TRIS (8.20 mmol). The reaction was heated to 70° C.for 0.5 hr. After cooling to room temperature, the solvent was removedin vacuum. The residue was sonicated in dichloromethane (10 mL) andconcentrated again. The resulting white solid was dried under the vacuumpump overnight. The crude material was crystallized by slurring thesolid residue in a 4:1 mixture of acetonitrile and methanol (5 mL). Themixture was stirred at room temperature for 24 hrs to give4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid TRIS salt as a white precipitate. Melting point (195.6° C.). ¹H NMR(400 MHz, DMSO): δ 7.92-7.80 (m, 2H), 7.78-7.64 (m, 1H), 7.41-7.19 (m,8H), 7.13-7.00 (m, 1H), 5.44 (s, 2H), 5.25-5.14 (m, 2H), 4.61-4.48 (m,2H), 4.18-4.03 (m, 3H), 3.39 (s, 7H).

In one embodiment,4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid (30.6 mmol) was added to 150 mL acetone to give a white suspension.To the suspension was gradually added 75 mL meglumine (34 mmol) watersolution. The resulting mixture was stirred at 50° C. for 4 h and thenat room temperature for 12 hrs to give4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid meglumine monohydrate salt as a white solid, which dehydrated atabout 71° C. as determined by DSC.

In another embodiment, further heating of the meglumine monohydrate saltat 80° C., 0% relative humidity in an oven for 30 min. gave a whitecrystalline solid having a melting point (Tm onset) of 167.5° C.Alternatively,4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid meglumine monohydrate salt (0.1 mmol) was added to 1 mL 1:1 (v/v)acetone/water. The resulting mixture was equilibrated at 50° C. for oneweek to give a white solid, which dehydrated at about 61° C. asdetermined by DSC.

In yet another embodiment, methanol (100 mL) was added to4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid (4.1 mmol) and meglumine base (4.1 mmol). The resulting suspensionwas refluxed at 80° C. for 24 hrs then cooled to room temperature togive4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid meglumine salt as a white solid. Melting point (180.6° C.).

The following compounds were prepared according to the proceduresdescribed in Example 14 using the appropriate intermediates.

Compound Structure Characterization Data 76

¹H NMR (400 MHz, DMSO): δ 7.74-7.66 (m, 1H), 7.61-7.54 (m, 1H),7.51-7.42 (m, 1H), 7.39-7.23 (m, 6H), 7.18-7.02 (m, 2H), 5.48-5.39 (m,2H), 5.22 (s, 2H), 4.59-4.48 (m, 2H), 4.18-4.06 (m, 3H). Mixture ofrotamers. MS (m/z): 506.2/508.2 (M + H)⁺ (chlorine isotope pattern). 77

¹H NMR (400 MHz, DMSO): δ 7.94-7.86 (m, 2H), 7.58-7.51 (m, 1H), 7.37 (m,3H), 7.19-7.02 (m, 5H), 5.40 (s, 2H), 5.33- 5.22 (m, 2H), 4.67-4.55 (m,2H), 4.15- 4.05 (m, 3H). Mixture of rotamers. MS (m/z): 490.2 (M + H)⁺.78

¹H NMR (400 MHz, DMSO): δ 7.93-7.85 (m, 2H), 7.71 (m, 1H), 7.34 (m, 3H),7.03 (m, 4H), 5.44 (s, 2H), 5.28 (m, 2H), 4.62 (m, 2H), 4.13 (s, 3H).Mixture of rotamers. MS (m/z): 524.2/526.2 (M + H)⁺ (chlorine isotopepattern). 79

¹H NMR (400 MHz, DMSO): δ 7.77-7.71 (m, 1H), 7.65-7.60 (m, 1H),7.60-7.52 (m, 1H), 7.43-7.24 (m, 6H), 7.19-7.11 (m, 1H), 7.10-7.03 (m,1H), 5.42-5.37 (m, 2H), 5.33-5.27 (m, 2H), 4.65-4.59 (m, 2H), 4.16-4.06(m, 3H). Mixture of rotamers. MS (m/z): 490.2 (M + H)⁺. 80

¹H NMR (400 MHz, DMSO): δ 7.94-7.88 (m, 2H), 7.73-7.69 (m, 1H),7.41-7.31 (m, 3H), 7.23 (s, 1H), 7.11-7.03 (m, 4H), 5.47-5.41 (m, 2H),5.28-5.17 (m, 2H), 4.62-4.51 (m, 2H), 4.17-4.07 (m, 3H), 2.28 (s, 3H).Mixture of rotamers. MS (m/z): 502.2/504.2 (M + H)⁺ (chlorine isotopepattern). 81

¹H NMR (400 MHz, DMSO): δ 7.95-7.84 (m, 2H), 7.71 (s, 1H), 7.44-7.27 (m,4H), 7.22-7.00 (m, 3H), 5.44 (s, 2H), 5.39- 5.31 (m, 2H), 4.74-4.60 (m,2H), 4.17- 4.05 (m, 3H). Mixture of rotamers. MS (m/z): 524.2/526.2 (M +H)⁺ (chlorine isotope pattern). 82

¹H NMR (400 MHz, DMSO): δ 7.95-7.84 (m, 2H), 7.71 (s, 1H), 7.44-7.27 (m,4H), 7.22-7.00 (m, 3H), 5.44 (s, 2H), 5.39- 5.31 (m, 2H), 4.74-4.60 (m,2H), 4.17- 4.05 (m, 3H), 2.28 (s, 3H). Mixture of rotamers. MS (m/z):520.2/522.2 (M + H)⁺ (chlorine isotope pattern). 83

¹H NMR (400 MHz, DMSO): δ 7.90 (m, 2H), 7.56 (m, 1H), 7.39 (m, 2H),7.20- 6.91 (m, 5H), 5.40 (s, 2H), 5.37-5.19 (m, 2H), 4.72-4.52 (m, 2H),4.12 (s, 3H). Mixture of rotamers. MS (m/z): 508.2 (M + H)⁺. 84

¹H NMR (400 MHz, DMSO): δ 7.77-7.68 (m, 2H), 7.65-7.59 (m, 1H),7.45-7.31 (m, 2H), 7.26-7.19 (m, 1H), 7.12-7.03 (m, 4H), 5.48-5.39 (m,2H), 5.34-5.18 (m, 2H), 4.67-4.46 (m, 2H), 4.21-3.90 (m, 3H), 2.27 (s,3H). Mixture of rotamers. MS (m/z): 520.2/522.2 (M + H)⁺ (chlorineisotope pattern). 85

¹H NMR (400 MHz, DMSO): δ 7.79-7.68 (m, 1H), 7.65-7.50 (m, 2H),7.45-7.36 (m, 1H), 7.27-7.02 (m, 6H), 5.43-5.34 (m, 2H), 5.34-5.21 (m,2H), 4.68-4.50 (m, 2H), 4.20-3.94 (m, 3H), 2.27 (s, 3H). Mixture ofrotamers. MS (m/z): 504.2 (M + H)⁺. 86

¹H NMR (400 MHz, DMSO): δ 11.22- 11.02 (m, 1H), 7.97-7.87 (m, 2H), 7.76-7.66 (m, 1H), 7.46-7.30 (m, 6H), 7.10- 6.99 (m, 9.8, 2H), 6.43-6.37 (m,1H), 5.51-5.41 (m, 2H), 5.31-5.16 (m, 2H), 4.66-4.54 (m, 2H), 4.22-4.05(m, 3H). Mixture of rotamers. MS (m/z): 527.2 (M + H)⁺. 87

¹H NMR (400 MHz, DMSO): δ 8.65-8.53 (m, 1H), 8.03-7.95 (m, 1H),7.90-7.78 (m, 1H), 7.74-7.68 (m, 1H), 7.43-7.31 (m, 2H), 7.19-7.04 (m,4H), 5.47-5.40 (m, 2H), 5.34-5.27 (m, 2H), 4.70-4.61 (m, 2H), 4.17-4.07(m, 3H). Mixture of rotamers. MS (m/z): 507.2 (M + H)⁺. 88

¹H NMR (400 MHz, DMSO): δ 8.50-8.44 (m, 1H), 8.43-8.32 (m, 1H),7.94-7.83 (m, 2H), 7.73-7.68 (m, 1H), 7.68-7.54 (m, 1H), 7.45-7.39 (m,1H), 7.39-7.30 (m, 2H), 7.10-7.02 (m, 1H), 5.48-5.24 (m, 4H), 4.73-4.59(m, 2H), 4.16-4.08 (m, 3H). Mixture of rotamers. MS (m/z): 507.1 (M +H)⁺. 89

¹H NMR (400 MHz, DMSO): δ 8.53-8.40 (m, 2H), 7.93-7.68 (m, 4H),7.46-7.30 (m, 3H), 7.09-7.02 (m, 1H), 5.48-5.22 (m, 4H), 4.74-4.57 (m,2H), 4.17-4.05 (m, 3H). Mixture of rotamers. MS (m/z): 523.2 (M + H)⁺.90

¹H NMR (400 MHz, CDCl3): δ 7.82 (dd, J = 8.1, 17.0 Hz, 1H), 7.70 (dd, J= 2.5, 6.6 Hz, 1H), 7.43-7.28 (m, 2H), 7.13 (ddd, J = 7.8, 22.1, 26.9Hz, 6H), 5.44 (d, J = 4.6 Hz, 2H), 5.28 (s, 2H), 4.61 (s, 2H), 4.12 (d,J = 15.7 Hz, 3H). MS (m/z): 524.2/526.2 (M + H)⁺ (chlorine isotopepattern). 91

¹H NMR (400 MHz, DMSO): δ 7.83 (dd, J = 7.9, 17.5 Hz, 1H), 7.71 (dd, J =2.5, 9.4 Hz, 1H), 7.32 (ddd, J = 4.7, 12.1, 20.5 Hz, 6H), 7.24-7.00 (m,3H), 5.44 (d, J = 7.0 Hz, 2H), 5.26 (d, J = 11.9 Hz, 2H), 4.59 (d, J =4.5 Hz, 2H), 4.13 (d, J = 22.5 Hz, 3H). Mixture of rotamers. MS (m/z):506.2/508.2 (M + H)⁺ (chlorine isotope pattern). 92

¹H NMR (400 MHz, DMSO): δ 7.92 (app d, J = 8.0 Hz, 2H), 7.72 (br s, 1H),7.46-7.22 (m, 5 H), 7.16-7.08 (m, 2H), 7.04 (dd, J = 1.3, 8.0 Hz, 1H),5.42 (br s, 2H), 5.32 (br s, 2H), 4.62-4.54 (m, 2H), 4.14-4.05 (m, 3H).Mixture of rotamers. MS (m/z): 506.1/ 508.1 (M + H)⁺ (chlorine isotopepattern). 93

MS (m/z): 524.1/526.1 (M + H)⁺ (chlorine isotope pattern). r.t = 1.70994

¹H NMR (400 MHz, DMSO): δ 12.03 (m, 1H), 7.68 (d, J = 7.5 Hz, 1H), 7.56(d, J = 8.3 Hz, 1H), 7.48 (dd, J = 1.8, 7.5 Hz, 1H), 7.39-7.32 (m, 2H),7.28-7.11 (m, 4H), 7.02 (dd, J = 1.9, 8.4 Hz, 1H), 5.33 (br s, 2H), 5.26(s, 2H), 4.72 (br s, 2H), 3.98 (s, 3H). MS (m/z): 508.1 (M + H)⁺. 95

MS (m/z): 506.1/508.1 (M + H)⁺ (chlorine isotope pattern); r.t = 1.80196

¹H NMR (400 MHz, DMSO): δ 12.33 (m, 1H), 7.83 (app t, J = 7.9 Hz, 2H),7.76 (d, J = 8.3 Hz, 1H), 7.48-7.42 (m, 3H), 7.24- 7.01 (m, 4H), 5.38(br s, 2H), 5.28 (s, 1H), 5.20 (s, 1H), 4.70 (br s, 2H), 4.06-4.00 (m,3H). Mixture of rotamers. MS (m/z): 524.1/ 526.1 (M + H)⁺ (chlorineisotope pattern). 97

MS (m/z): 524.1/526.1 (M + H)⁺ (chlorine isotope pattern); r.t = 1.75198

¹H NMR (400 MHz, d₄-MeOH): δ 9.01 (br s, 1H), 8.42 (app t, J = 8.2 Hz,1H), 7.66- 7.58 (m, 2H), 7.28-6.89 (m, 6H), 5.37-5.31 (m, 4H), 4.74 (brs, 1H), 4.68 (br s, 1H), 4.04-3.98 (m, 3H, NMe rotamers visible).Mixture of rotamers. MS (m/z): 507.1/ 509.1 (M + H)⁺ (chlorine isotopepattern). 99

¹H NMR (400 MHz, DMSO): δ 12.54 (br s, 1H), 7.78 (d, J = 7.6 Hz, 1H),7.72-7.64 (m, 1H), 7.54 (app dd, J = 1.5 Hz, 7.8 Hz, 1H), 7.39-7.22 (m,2H), 7.19-7.08 (m, 3H), 6.98 (app d, J = 8.0 Hz, 1H), 5.38 (app d, J =10 Hz, 2H), 5.24 (br s, 2H), 4.73 (br s, 1H), 4.64 (br s, 1H), 4.13-3.97(m, 3H), 2.45 (app d, J = 11.2 Hz, 3H). Mixture of rotamers. MS (m/z):521.1/523.1 (M + H)⁺ (chlorine isotope pattern).

Example 154-Fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid (Compound 100)

Step 1

HATU (1.2 mmol) was added to a mixture of8-fluoro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid(I-5) (1.1 mmol), Hunigs Base (2.4 mmol) and DMF (5 mL). A solution ofmethyl 4-fluoro-3-((2-((3-fluorobenzyl)amino)ethyl)amino)benzoate (I-46)(1.1 mmol) in DMF (3 mL) was added and the reaction mixture stirred atroom temperature for 4 hrs. The mixture was diluted with water and ethylacetate. The layers were separated and the aqueous phase was washed withethyl acetate. The combined organic extracts were washed with water,brine, and then dried over MgSO₄. The material was purified bychromatography (silica, 0-60% ethyl acetate/hexanes) to give methyl4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoate(100-Int).

Step 2

1N KOH (4 mmol, 5 equiv.) was added to a solution of methyl4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoate(100-Int) (0.80 mmol) in THF (4 mL) and MeOH (2 mL) and the solution wasstirred at 50° C. for 2 hrs. The solvent was removed and the cruderesidue was diluted with water. The aqueous solution was acidified withacetic acid (pH ˜5) to give4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid as a white precipitate. ¹H NMR (400 MHz, DMSO): δ 7.61-7.26 (m,2H), 7.20-6.82 (m, 9H), 6.11-5.81 (m, 1H), 5.43-4.70 (m, 4H), 4.15-3.94(m, 4H), 3.53 (t, J=6.6 Hz, 1H), 3.44-3.39 (m, 2H). MS (m/z): 537.2(M+H)⁺; r.t.=1.618; Elemental Analysis: calcd. for 0.50,C₂₈H₂₂F₃N₄O*0.52 H₂O: C, 61.65; H, 4.43; N, 10.45); found: C: 61.75; H:4.21; N: 10.31.

4-Fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid (7.5 mmol) added to a solution of TRIS (7.5 mmol) in MeOH (400 mL).The reaction was stirred at 60° C. for 30 mins. After cooling to roomtemperature, the solvent was removed and the crude material wasazeotroped with dichloromethane (2×). The resulting glassy solid wasslurred in ethyl acetate (200 mL). The slurry was stirred at roomtemperature for 24 hrs. The resulting solid was collected by filtrationto give4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid TRIS salt as a solid. Melting point (160° C.). ¹H NMR (600 MHz,DMSO): δ 7.62-6.70 (m, 10H), 5-90-5.80 (m, 1H), 5.45-5.15 (m, 2H),4.75-4.55 (m, 2H), 4.16-3.5 (m, 4H), 3.41-3.39 (m, 1H), 3.30-3.25 (m,13H). Anal. Calcd for C₃₂H₃₄F₃N₅O₇*1.3 H₂O: C, 56.36; H, 5.42; N, 10.27*1 H₂O Found: C, 56.25; H, 5.29; N, 10.32.

A solution of L-arginine (0.20 mmol) in deionized water (3 mL) was addedto a suspension4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoicacid (0.20 mmol) in MeOH (12 mL). The mixture was stirred at 70° C. for0.5 hr and then stirred for 2 hrs at room temperature. The solvent wasremoved under vacuum and the crude material was crystallized by slurringthe solid residue in acetonitrile (5 mL). The mixture was stirred atroom temperature for additional 24 hrs to give4-fluoro-3-((2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)benzoic acid L-arginine salt as a white precipitate. Melting point (161°C.).

Example 163-((2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid (Compound 101)

3-((2-(8-chloro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)ethyl)amino)-4-fluorobenzoicacid was prepared according to the procedure described in Example 13using the appropriate intermediates. MS (m/z): 553.2/555.2 (M+H)⁺(chlorine isotope pattern); r.t.=1.871.

Biological Assays Human GST-FXR LBD Co-Activator Interaction Assay

The FXR HTRF assay is a biochemical assay measuring the interactionbetween FXR and a coactivator protein (SRC1). The ligand-inducedinteraction with a coactivator protein is a critical step intranscriptional activation by FXR. Thus, this is an assay designed tomeasure FXR agonist activity of the test compounds.

Recombinant human Farnesoid X Receptor (FXR) ligand binding domain(amino acids 193-472) fused to glutathione S-transferase (GST) purifiedprotein (GST-FXR LBD) was purchased (Invitrogen). The ligand-dependentinteraction between GST-FXR LBD and a peptide derived from SteroidReceptor Coactivator-1 (SRC-1) was monitored by Fluorescence ResonanceEnergy Transfer (FRET). GST-FXR LBD was mixed with a biotin-labeledSRC-1 peptide (Biotin-CPSSHSSLTERHKILHRLLQEG-SPS-CONH2 (SEQ ID NO:1),American Peptide) in assay buffer (50 mM Tris.HCl, pH 7.4, 50 mM NaCl, 1mM TCEP and 0.2% bovine serum albumen) and plated in 384 black Proxiplates (Greiner Bio-One). Test compounds (in DMSO solution) anddetection reagents (anti-GST-Cryptate labeled antibody andStreptavidin-XL665 conjugate; CisBio) were added in assay buffercontaining 50 mM KF. Plates are incubated at room temperature in thedark for 2.5 hrs before reading on an Envision (Perkin Elmer) at 665 nmand 590 nm. The HTRF assay results were calculated from the 665 nm/590nm ratio (ratio=(A665 nm/A590 nm)×10⁴) and expressed in Delta F%=(Ratiosample−Rationegative)/Rationegative×100.

A negative control (without Streptavidin-XL665) was run with each assayand represented the background fluorescence. A reference FXR agonist,(E)-3-(2-chloro-4-((3-(2,6-dichlorophenyl)-5-isopropylisoxazol-4-yl)methoxy)styryl)benzoicacid (Compound GW4064), was included in each experiment as positivecontrol. The efficacy of each test compound was compared to that ofGW4064. At each concentration, the relative activity of the testcompound was expressed as Response%=(R_(sample)−R_(DMSO))/(R_(positive)−R_(DMSO)), where R_(sample) is theHTRF response (expressed in Delta F %) for the test compound,R_(positive) is the maximal response for GW4064 at saturatingconcentrations, and R_(DMSO) is the response for DMSO control. The EC₅₀values were calculated using GraphPad Prism (GraphPad Software) usingnon-linear regression curve fit (log(agonist) vs. response−variableslope (four parameters)).

Table 1 summarizes EC₅₀ values for the compounds of the invention inhuman GST-FXR LBD co-activator interaction assay.

TABLE 1 Compound FXR coactivator interaction Number assay (HTRF) (μM) 10.0012 2 0.0006 3 0.00051 4 0.00075 5 0.00080 6 0.0024 7 0.0010 8 0.00339 0.0036 10 0.0017 11 0.0033 12 0.0054 13 0.00080 14 0.00084 15 0.0022216 0.0034 17 0.0010 18 0.0004 19 0.0005 20 0.0005 21 n.d. 22 n.d. 23n.d. 24 0.0032 25 0.0007 26 n.d. 27 0.0014 28 0.00038 29 0.00067 300.00060 31 0.00047 32 0.00074 33 0.0015 34 0.0021 35 0.00077 36 0.033 370.024 38 0.0026 39 0.0059 40 0.0086 41 0.0040 42 0.0025 43 0.018 440.011 45 0.015 46 0.0032 47 0.00053 48 0.061 49 0.017 50 0.011 51 0.009852 0.0026 53 0.017 54 0.021 55 0.013 56 0.040 57 0.030 58 0.022 59 0.02860 n.d. 61 0.089 62 0.074 63 0.0060 64 0.00060 65 0.007 66 n.d. 67 n.d.68 n.d. 69 0.025 70 0.081 71 0.022 72 0.0026 73 0.020 74 0.055 75 0.00676 0.0096 77 0.0079 78 0.0083 79 0.022 80 0.016 81 0.021 82 0.012 830.014 84 0.018 85 0.035 86 0.012 87 0.028 88 0.015 89 n.d. 90 0.005 910.0065 92 0.014 93 0.013 94 0.032 95 0.0035 96 0.0036 97 0.017 98 n.d.99 0.029 100 0.024 101 0.0055 n.d. = not determined

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference for allpurposes.

We claim:
 1. A compound selected from the group consisting of:4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid L-arginine salt;4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid sodium salt;4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid L-arginine salt;4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid L-lysine salt;4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid TRIS salt;4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid meglumine monohydrate; and4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid meglumine salt.
 2. A pharmaceutical composition comprising atherapeutically effective amount of a compound of claim 1 or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier.
 3. A combination comprising a therapeuticallyeffective amount of a compound of claim 1 or a pharmaceuticallyacceptable salt thereof, and a second therapeutic agent.
 4. A method fortreating a condition mediated by farnesoid X receptors (FXR) in asubject suffering therefrom, comprising administering to the subject atherapeutically effective amount of a compound of claim 1 or apharmaceutically acceptable salt thereof, and optionally in combinationwith a second therapeutic agent.
 5. The method of claim 4, wherein saidcondition mediated by FXR is a liver disease or a gastrointestinaldisease.
 6. The method of claim 5, wherein said condition mediated byFXR is a liver disease selected from intrahepatic cholestasis,estrogen-induced cholestasis, drug-induced cholestasis, cholestasis ofpregnancy, parenteral nutrition-associated cholestasis, progressivefamilial cholestasis (PFIC), Alagille syndrome, primary biliarycirrhosis (PBC), primary sclerosing cholangitis, ductopenic livertransplant rejection, liver transplant associated graft versus hostdisease, cystic fibrosis liver disease, non-alcoholic fatty liverdisease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic liverdisease, and parenteral nutrition-associated liver disease.
 7. Themethod of claim 6, wherein said condition mediated by FXR isnon-alcoholic fatty liver disease (NAFLD) or non-alcoholicsteatohepatitis (NASH).
 8. The method of claim 5, wherein said conditionmediated by FXR is a gastrointestinal disease selected from primary bileacid diarrhea, secondary bile acid diarrhea, bile reflux gastritis, andinflammatory bowel disease.
 9. The method of claim 4, wherein saidcompound is4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid L-arginine salt.
 10. The method of claim 4, wherein said compoundis4-fluoro-3-(2-(8-fluoro-N-(2-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid sodium salt.
 11. The method of claim 4, wherein said compound is4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid L-arginine salt.
 12. The method of claim 4, wherein said compoundis4-fluoro-3-(2-(8-fluoro-N-(3-fluorobenzyl)-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)acetamido)benzoicacid L-lysine salt.
 13. The method of claim 4, wherein said compound is4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid TRIS salt.
 14. The method of claim 4, wherein said compound is4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid meglumine monohydrate.
 15. The method of claim 4, wherein saidcompound is4-((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamido)methyl)benzoicacid meglumine salt.